bluuebunny/crossref_metadata_2025 huggingface dataset:
GPR18 Distribution and Expression in Human Placenta
Authors: Yuil‐Valdes Ana , Mukherjee Maheswari , Anderson‐Berry Ann , Hanson Corrine K. , Berry Elizabeth , Nordgren Tara
Published: June 2021

Background

Optimally functioning inflammation resolution systems during pregnancy are essential in regulating a series of complex processes that could degenerate into persistent inflammation and result in poor maternal‐child outcomes. Infants developing in a pro‐inflammatory intrauterine environment are at significantly higher risk for severe complications than infants born without pregnancy inflammatory risk factors.

Specialized pro‐resolving lipid mediators (SPM) are chemical structures derived from polyunsaturated fatty acids. Resolution of acute inflammation is an active process regulated by SPM, including resolvin D2 (RvD2), a potent docosahexaenoic acid‐derived resolution‐phase mediator, which activates a cell surface G protein–coupled receptor, GPR18/RvD2.

Preliminary data from our group indicate that the SPM RvD2 is significantly increased in maternal plasma taken at the time of delivery compared to cord blood plasma (3,450 vs. 689 pg/mL for maternal vs. cord, respectively, p<0.001), with further increases in the maternal plasma of cases with adverse maternal‐fetal outcomes, including NICU admission. Based on this data, we hypothesize that the placenta is a target site of action of SPM during perinatal growth and development. We identified the expression of the RvD2 receptor, GPR18 by immunostain on decidual cells, smooth muscle of the maternal and fetal arteries and syncytiotrophoblast, cytotrophoblast and macrophages of chorionic villi. Since the decidual cells showed significant expression of GRP18 therefore, the objective of this study was to analyze and categorize the different levels of staining intensity in this cell type.

Methods

Ten cross‐sections of placental tissues containing both maternal and fetal sides were stained for SPM‐binding receptor, GPR18 immunostaining, and digitized at 40× magnification using VENTANA iScan HT scanner. Our novel approach including Definiens Tissue Studio® software was used to quantify the cellular analyses of decidual cells in these digitized images that quantify low, medium, and high expressions for GPR18.

Results

The majority of decidual cells showed medium or high GPR18 membranous and cytoplasmic expression (Figure 1). 20% of decidua cells had no areas of low expression. Seven out of ten samples had areas of medium expression that were greater than 50%. Areas of low expression ranged from 0–31%, while areas of medium expression ranged from 11–81%, and high expression ranged from 5–75%.

Conclusions

The decidual cells of the placenta have medium/high membranous and cytoplasmic expression of GPR18. SPM may be an important component of the inflammation physiology in the placental tissue. The expression of GPR18 may play a protective role in pregnancy and perinatal disease.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Profiling apolipoproteins in disease
Authors: Dreskin Elizabeth Jordan , Brampton Chris , Houser Brett , Hendricksen Amy
Published: June 2021

Detecting changes in biomarker levels or even the presence of biomarkers in limited samples can be difficult with traditional ELISA techniques. Multiplexing magnetic bead based assays not only affords much higher sensitivity, but also allows for sample conservation by testing for multiple analytes in a single well. In this study, we used an apolipoprotein multiplex panel to screen for some of the traditional apolipoproteins within the statin pathway (ApoA1, ApoA2, ApoB, ApoC1, ApoC3, and ApoD , along with additional targets important for cardiovascular disease, neurobiology, cancer, and inflammation research; Apo E, Apo H, Clusterin (Apo J) and C‐Reactive Protein (CRP). Purchased serum samples representing several different diseases, including atherosclerosis, Diabetes Mellitus Type I, sepsis, Alzheimer's, and Traumatic Brain Injury, were diluted 1:50,000. With the exception of CRP levels in some sepsis samples, all readings were within range. We conclude that this technology, and specifically this Apolipoprotein 10‐plex, can be applied to many areas of disease research, and requires a minimum of sample of serum in order to generate robust data.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RELATION ASYMMETRY FACIAL AND SYMMETRY FACIAL vs TIMES AND FIXATIONS TO BUILD A DIAGNOSIS IN A HISTOPATHOLOGY TASK
Authors: MONDRAGON EDWARD JAVIER ACERO
Published: June 2021

The asymmetry facial is a signal morphometric that in the development embryonal increase by causes endogens how mutations, homozygosity, inbreeding, and causes exogens how minors epigenetic adaptations during environmental stresses maternal. During task of observation the perceptual load has individual differences relatively high by fixations over shape, size, brightness and color and the experience of observation, but don't exist references about its relations with asymmetry facial when is accepted in biology that asymmetry impose different performances functionals

Materials and Methods

Second‐year medical students 8 females (□ = 18.4 years) and 6 males (□ = 18.9 years) were calculated their facial asymmetry being n = 4 symmetrical and n = 9 asymmetric; In time of free interaction the entire population was challenged to the definitive histopathological diagnosis of basal cell carcinoma from digital images, measuring both duration and quantity of fixations.

RESULTS

Between symmetric individuals and histopathological diagnosis, there was a negative correlation coefficient r = −0.9411 between the duration of their fixations (average 310 ms, DS = 40 ms) and the number of their fixations (mean 103, DS = 16 ), Which has significant differences in the performance of asymmetric individuals with a less negative correlation coefficient r = −0.743 between the duration of fixations (average 237 ms, DS = 28 ms) and the number of fixations (average 144, SD = fifty).

Conclusion

Symmetric individuals do less fixations and last longer in them when they make histopathological diagnosis of basal cell carcinoma with respect to asymmetric individuals who make more fixations and last less in each one.
Agnostic Pathology of Molecular Biomarkers in Smell Loss (Hyposmia)
Authors: Henkin Robert , Abdelmeguid Mona
Published: June 2021

There are multiple pathologies which biochemically initiate and perpetuate smell loss (hyposmia). These pathologies are non‐neoplastic diseases and include viral‐type infections, allergic rhinitis, traumatic head injuries, inhalational anesthesia and several other pathologies. We have attempted to evaluate specific biochemical parameters, manifested as biomarkers, which identify the pathology underlying these various causes of hyposmia. While each of these pathologies reflects widely different initiating events to cause hyposmia the biomarkers underlying these events are similar in each pathology. These biomarkers are found in nasal mucus, the biological fluid which maintains olfactory receptors. These biomarkers are decreased nasal mucus concentrations of cAMP, cGMP and sonic hedgehog. The mechanisms for these changes are similar across each of these pathologies since these biomarkers reflect diminished activity of these moieties which act as growth or transcription factors which stimulate stem cells in the olfactory epithelium to initiate growth and perpetuation of olfactory epithelial receptor cells. Similarities in these growth factor abnormalities across these multiple initiating pathological events are the basis for this agnostic determination of the biomarkers which induce hyposmia. Indeed, correction of these biochemical abnormalities with treatment with the phosphodiesterase inhibitor theophylline increases nasal mucus levels of each of these biomarkers in each pathology which thereby improves hyposmia in each pathological entity and establishes the agnostic character of these biomarkers.

Support or Funding Information

NA

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Skeletal Muscle Produces Acute Phase Proteins in Response to Polymicrobial Sepsis
Authors: Robinson Gerard P. , Mattingly Alex J. , Garcia Christian K. , Murray Kevin O. , Clanton Thomas L. , Laitano Orlando
Published: June 2021

Skeletal muscle has been implicated as an endocrine organ, as observed by the secretion of cytokines in response to multiple stimuli such as exercise, heat stress and infection. However, whether skeletal muscles can produce acute phase proteins (APPs) in response to polymicrobial sepsis is controversial. This is relevant since APPs play an important role in the transition from innate to adaptive immunity and, thus far, the liver has been considered the primary source of these proteins. Our aim was to determine whether skeletal muscles are capable of producing APPs in response to polymicrobial sepsis.

METHODS

We injected (IP) 37 female C57BL/6J mice with cecal slurry (1.6 g/ml) and harvested the soleus, gastrocnemius and livers at 6, 12, and 24 h after the injection. Respective muscles and livers from non‐septic mice were used as controls. Specimens were then homogenized and analyzed for protein content. Immunoblot assays were then conducted using polyclonal antibodies for, serum amyloid a1 (SAA1), fibrinogen, lipocalin, and c‐reactive protein (CRP). For quantification, membranes were scanned for fluorescence intensity and normalized for total protein.

RESULTS

We observed elevated expression of fibrinogen, lipocalin and SAA1 in the livers of septic mice across all time points in comparison to non‐septic mice. We observed an increased signal for SAA1 and fibrinogen in skeletal muscle of septic mice. Notably, the signal intensity was higher in soleus in comparison to gastrocnemius muscle, which suggests a muscle‐specific response. We did not observe any signal for lipocalin, regardless of the muscle type. We also detected the presence of CRP in the soleus muscles of septic mice. Our results suggest that the cecal slurry model of septic shock induces an acute phase response in both liver and skeletal muscle. Though skeletal muscles produce a smaller spectrum of known APPs, these may contribute to the transition from innate to adaptive immunity, and the overall response to severe bacterial infection.

Support or Funding Information

Supported by NIH RO1GM118895‐01 and the BK and Betty Stevens Endowment.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of Potential Broad‐Spectrum Peptidomimetic Antivirals Active Against Zika Virus
Authors: Khachatoorian Ronik , Buzzanco Anthony S. , Waring Alan , Riahi Rana , Jin Daphne , Dasgupta Asim , Arumugaswami Vaithilingaraja , Ruchala Piotr , French Samuel W.
Published: June 2021

Zika virus (ZIKV) has recently been the cause of major epidemics in South American countries especially Brazil. ZIKV has been identified as the cause of microcephaly in the fetuses of pregnant women and the Guillain‐Barre Syndrome, and to date, no therapeutics or vaccines are available against ZIKV infection. Previously, we reported the anti‐ZIKV activity of Heat Shock Protein (HSP) 70‐Interacting Peptide (HIP), a cell‐permeable hairpin peptide derived from the domain I of nonstructural protein 5A of hepatitis C virus (HCV). These results implicate the HSP70 family members in the pathogenesis of ZIKV, identify the HSP70 family proteins as potential host therapeutic targets against ZIKV, and present the HIP as a potential therapeutic agent with significant anti‐ZIKV activity. In this study, we performed rigorous in silico analyses to identify potential peptidomimetic compounds that mimic the binding of the HIP to HSP70. Through particle swarm optimization via SwarmDock, binding solutions were generated from crystal structures of HSP70 nucleotide binding domain (NBD) and NS5A Domain I. The highest‐ranked residue interactions were mapped through visualization software, which identified a common binding region at a pocket at the C‐terminus. Subsequent modeling at this pocket was used to generate a HIP‐binding model, independently confirmed through docking of HIP to HSP70‐NBD in AutoDock Vina. Virtual high‐throughput screening of a diverse compound library against the binding pocket identified putative leads. We are currently assessing the antiviral activity of the lead compounds in our cell culture model of ZIKV infection. Subsequently, the best leads will be optimized to achieve greater antiviral activity. As HIP also blocks HCV, the targeting of HSP70 may represent a broad‐spectrum approach for future antivirals. Targeting ZIKV and other Flaviviruses through host proteins provides a therapeutic target that avoids selection of resistant viral particles.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Extracellular vesicles secreted by Trypanosoma cruzi: relationship with polyunsaturated fatty acids in the modulation of infection
Authors: Lovo‐Martins Maria Isabel , Malvezi Aparecida Donizette , Zanluqui Nágela Ghabdan , Lucchetti Bruno Fernando Cruz , Suzukawa Helena Tiemi , Tatakihara Vera Lucia Hideko , Goldenberg Samuel , Fritsche Kevin , Wowk Pryscilla Fanini , Pinge‐Filho Phileno Isabel
Published: June 2021

Fish oil (FO) containing docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) has been reported to exert beneficial health effects on the host in the early course of Trypanosoma cruzi infection. T. cruzi virulence factors include molecules expressed on the cell surface as well as those secreted or shed into the extracellular medium. Extracellular vesicles shed by T. cruzi (EV) promote infection susceptibility of mammalian cells. We examined whether fish oil supplementation could prevent the effects of EV in the early course of T. cruzi infection. We show that the inoculation of EV from Y strain of T. cruzi (EV Y), in C57BL/6 mice prior to T. cruzi infection resulted in increased parasitemia, increased cardiac parasitism and decreased nitric oxide (NO) production. Supplementation of the animals with FO prevented the increase of parasitemia in animals pre‐treated with EV Y and impairs the action of EV Y on NO production by spleen cells stimulated with Tc‐Ag. These results suggest that EV Y modulate the host response in favor of the parasite and that the presence of n‐3 PUFAs interfere in the action exerted by EV Y. This is the first description of EV from T. cruzi with n‐3 PUFAS‐dependent effects.

Support or Funding Information

This work was supported by CNPq (Edital Universal 14‐2014), and Fundação Araucária (grant No. 419‐2009). MILM, NGZ, BFCL and HTS, received scholarships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). PPF received research fellowship from CNPq.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Contribution of Zika Virus Envelop Protein‐Induced Autoantibodies in Neuropathy
Authors: Lai Yen‐Chung
Published: June 2021

Zika virus (ZIKV) belongs to flaviviridae family, which is transmitted by the same vectors as dengue virus (DENV). However, unlike DENV, ZIKV infection can cause neurological manifestations such as fetal neurodevelopmental retardation and microcephaly in pregnant women and Guillain‐Barré syndrome in adult. Although autoimmunity is found to contribute to neuropathy in several diseases, the correlation of autoimmunity in neuron pathogenesis of ZIKV is still unclear. According to previous study, ZIKV polypeptides share antigenic similarities with neuron cells in predicted database. To investigate whether ZIKV‐elicited antibodies (Abs) could cross react to neuron cells, we immunized mice with recombinant envelop (E) protein and nonstructural protein 1(NS1) of ZIKV and DENV to generate Abs. Compare to DENV E protein and NS1, we found that Abs against ZIKV E protein could cross react to human neuronal cell, SK‐N‐SH cell line in a dose dependent manner. In addition, the binding of Abs against ZIKV E protein could induce caspase 3 activation and pro‐apoptosis gene, Bax, transcription in vitro. Furthermore, we demonstrated that passive transfer of Abs against ZIKV E protein in early embryonic phase of pregnant mice could lead to low fertility. Taken together, we suggested that ZIKV E protein could elicit autoantibodies that contribute to neuropathy.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Group A Streptococcus (GAS) Inhibits the Oxidative Burst Response in a Human Macrophage‐like Cell Line By Preventing the Formation of NADPH Oxidase Complex
Authors: Shiosaki Jessica , Yuen Isabelle , McLain Maya , Okumura Cheryl
Published: June 2021

Group A Streptococcus (GAS) is the cause of millions of infections worldwide each year, many of which progress to severe invasive infections such as sepsis, necrotizing fasciitis, and streptococcal toxic shock syndrome. Cells of the human innate immune system typically initiate the oxidative burst response to form reactive oxygen species (ROS) as a killing mechanism in response to pathogens. Other bacterial species including Lactococcus lactis and Staphylococcus aureus engage defense mechanisms such as catalase or pigments to detoxify these ROS. However, GAS lacks such detoxifying enzymes, and its survival in macrophages suggests its use of an alternative mechanism to avoid ROS‐mediated killing. When we infected THP‐1 macrophages with GAS, infected macrophages displayed significantly decreased peroxide production when compared to infection with S. aureus, L. lactis, and Group B Streptococcus. As the NADPH Oxidase (NOX2) complex is responsible for initiating the oxidative burst response and the generation of ROS such as peroxide, these data suggest that GAS may be impairing the production of ROS by either preventing the assembly of the NOX2 complex or its activation. The NOX2 complex forms on the surface of the phagosomal membrane and consists of two membrane proteins, gp91 and p22, which assemble with three cytosolic proteins, p47, p40, and p67. To assess whether GAS prevents the assembly of the NOX2 complex, we performed cell fractionation to isolate phagosomes of GAS and L. lactis‐infected macrophages. Western blot and mass spectrometry data indicate that NOX2 complex formation is indeed impaired in GAS‐infected cells. We are also currently performing immunoprecipitation experiments using infected THP‐1 cell lysates and antibodies for the p22 membrane subunit to look for NOX2 complex formation. Thus, these data will determine if GAS impairs the production of harmful ROS in macrophages by preventing the initial assembly of the NOX2 complex, ultimately allowing their persistence in an innate immune cell designed to kill them.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of the Arginine Deiminase Pathway in the Survival of Group A Streptococcus in the Phagolysosome
Authors: Chen Shuyi , Zhang Robert , Snipper Joshua , Okumura Cheryl
Published: June 2021

Group A Streptococcus (GAS) is a Gram positive bacterial species responsible for infections ranging from localized to invasive and lead to more than 500,000 deaths per year. When macrophages are infected by GAS, the cell actively brings bacteria into its phagosome. The phagosome fuses with lysosome, which becomes acidified and activates proteolytic enzymes to subsequently eliminate bacteria. We have developed a GAS strain expressing a pH‐sensitive green fluorescent protein and found that the protein was successfully degraded at a pH lower than 6. However, the fluorescent signal is not degraded when GAS is in macrophage phagolysosomes, indicating that GAS prevents lysosome acidification, enhancing its intracellular survival. We are currently examining the role of Arc enzymes that are involved in the arginine deiminase (ADI) pathway in preventing lysosomal acidification. GAS uses ADI pathway to catabolize hosts' arginine and convert it to ammonia and ATP, which could act as an acid buffer in the lysosome. In the ADI pathway, the ArcC protein is a carbamate kinase, a necessary protein in producing ammonia. ArcD encodes the anti‐porter of arginine and ornithine, and is important to bring in substrate and export by‐products. Using a cell killing assay, we found that GAS mutant strains lacking ArcC and ArcD proteins (ΔArcC and ΔArcD) survived less well upon infection of THP‐1 macrophage cells. Immunofluorescence data indicates all three strains traffic normally through the phagocytic pathway. We are currently assessing whether the ΔArcC and ΔArcD strains prevent phagolysosomal acidification using the pH‐sensitive green fluorescent protein as a probe. A better understanding on the crucial enzymes that prevent lysosome acidification provides insights for developing drug targets and partially reveals the mechanism that enhances GAS intracellular survival.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Environmental Enrichment on Relapse to Methamphetamine Seeking
Authors: Althobaiti Yusuf S.
Published: June 2021

Relapse to drug use is the most critical challenge in treating addiction. Therefore, preventing relapse is the most important approach in managing addictive disorder. Psychostimulant use is a critical health issue in Saudi Arabia and worldwide. Of note, methamphetamine (METH) is one of the most commonly used psychostimulant worldwide. In this study, we investigated the relapse to METH seeking using conditioned place preference (CPP) in animal model. The CPP apparatus consists of two conditioning chambers which are distinguished by both visual and tactile cues. Of note, evidence showed the positive effects of environmental enrichment on neurogenesis and neurochemical measures. Surprisingly, there is no previous study investigated the role of environmental enrichment in relapse to METH seeking. Here, we tested the effects of environmental enrichment on the relapse to METH seeking in rat model of drug addiction. The CPP experiment consists of four distinct phases; the habituation, acquisition, extinction, and reinstatement phase. Rats were allowed to explore the entire CPP apparatus without any treatment during the habituation phase. Each animal then received either saline or METH (1 mg/kg, i.p.) for 8 days during the acquisition phase, followed by housing in either isolated or enriched environment for 30 days during the extinction phase. Finally, rats were tested for reinstatement produced by i.p. injections of METH (1 mg/kg, i.p.). METH treatment during acquisition phase has significantly increased time spent in METH‐paired chamber as compared to saline‐paired chamber. No significant changes in time spent in either saline‐ or METH‐paired chambers following the extinction phase. A priming dose of METH significantly increased time spent in METH‐paired chamber as compared to saline‐paired chamber in animals that were housed in isolated environment. Interestingly, enriched environment blocked the reinstatement effect of METH. This indicates the important role of environmental enrichment as a treatment strategy in drug dependence.

Support or Funding Information

This work was supported by Award Number 1‐438‐6013 from the Deanship of Graduate Studies and Research, Taif University, SA.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Estrogens are Necessary for Expression of Cocaine Seeking in Male Rats
Authors: Acevedo Daniel Alejandro , Alvarado John , Torres Lilith , Santos‐Vera Bermary , Mueller Devin
Published: June 2021

It has been shown that, in female rats, estrogen is needed for the extinction of the cocaine‐seeking behavior. In this study, we are looking for the reduction of the cocaine‐seeking behavior by administrating Fadroloze (FAD) to inhibit the estrogens in male rats. We hypothesize that inhibiting estrogens in males, will show reduced expression, impaired extinction and reduced reinstatement in the cocaine‐seeking. To study the expression, extinction and reinstatement we performed the CPP paradigm. During conditioning animals were injected with cocaine 10mg/kg and confined to one side of the chamber for 20 min, or injected with saline and confined to the other side of the chamber for 20 min. In the extinction sessions, the animals are divided in two groups, one group received Fadrozole and the other received vehicle (saline) daily for 8 days. In the extinction CPP test the animals have access to both chambers for 30 min in daily sessions. The group that was given FAD show a significant reduction in the cocaine seeking compared with the vehicle group. These findings show that estrogen is needed for the cocaine‐seeking behavior and may be important in the extinction of the cocaine seeking in males. Learning how estrogen affects the cocaine seeking, extinction and reinstatement.

Support or Funding Information

This study was supported by UPR‐PRISE NIH R2GM096955, and by National Institutes of Health R01 DA038042.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impulsivity induced neuronal differentiation
Authors: Peyton Lee R. , Oliveros Alredo , Starski Phillip A. , Choi Doo‐Sup
Published: June 2021

Alcohol use disorder (AUD) represents an extensive global issue for disease and premature death. One facet of AUD is impulsivity, which is defined as a predisposition toward rapid, unplanned reactions to internal or external stimuli with decreased regard to the negative consequences of these reactions. Due to the potentially harmful consequences associated with impulsive decision making, impulsivity is considered an endophenotype for developing addictive behaviors. Interestingly, the dorsal hippocampus (dHip) has been revealed as an area of interest in addictive behaviors. The dHip is an important brain region involved in regulating behavioral adaptability and learning. Aberrant dHip function has been shown to contribute to impulsive and addictive behaviors. Differential reinforcement of low rates of behavior (DRL) schedules have demonstrated that the hippocampus is implicated in reward acquisition and impulsivity in humans and rodent models. Reward‐seeking induced impulsivity is thought to be driven via contextual recollection of drug‐induced euphoria to environmental cues, a process known to be regulated by the dHip. Our lab has previously demonstrated that adenosine A2A receptor and ERK‐driven impulsive behavior potentiates hippocampal neuroblast proliferation. However, the fate of these precursor cells and the precise role of ERK signaling in conjunction with impulsive reward‐seeking remain unknown. Here, we demonstrate that DRL‐mediated impulsive reward‐seeking with the 2‐choice reaction time task (2‐CSRTT) increase the number of BrdU labeled cells in the dHip dentate gyrus. Our results also show a significant increase in colocalization of BrdU+ and the neuronal marker NeuN+ in impulsive animals compared to untrained controls. These results suggest that increased impulsivity may drive neuronal differentiation indicating potential regulation of the neurogenic process.

Support or Funding Information

This work was supported by the Samuel C. Johnson for Genomics of Addiction Program at Mayo Clinic, the Ulm Foundation, the Golby Foundation, the Sands Family, David Lehr Research Award from American Society for Pharmacology and Experimental Therapeutics, and National Institute on Alcohol Abuse and Alcoholism (AA018799).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex Differences in Oxytocin Effects on CeA GABAergic Signaling in Alcohol‐Dependent and Naive Rats
Authors: Kirson Dean , Steinman Michael Q. , Roberto Marisa
Published: June 2021

Alcohol use disorders (AUDs) are significant public health concerns, characterized by compulsive seeking and consumption of alcohol. AUDs have been linked to disruption of normal functioning of brain stress systems, as the transition to dependence recruits these systems leading to the negative affective states seen in withdrawal; the relief of which drives further drinking. The central nucleus of the amygdala (CeA) functions as a neuropeptidergic hub of stress and anxiety processing, and GABAergic signaling within the CeA is involved in the regulation of alcohol consumption. Oxytocin is a stress‐related neuropeptide that has previously been shown to alter GABAergic signaling within the CeA of rodents. Oxytocin also affects alcohol addiction, dependence, and withdrawal, as administration of oxytocin decreases drinking and blocks alcohol withdrawal symptoms in humans and rodents. However, characterization of the electrophysiological effects of oxytocin and alcohol on GABAergic signaling in the CeA have not yet been reported. In this study, we used whole cell patch clamp electrophysiological recordings to examine the effects of oxytocin and alcohol on CeA GABAergic signaling in both alcohol naïve and chronic intermittent alcohol vapor treated (alcohol dependent) Sprague‐Dawley male and female rats. We also used immunohistochemical staining to identify alcohol dependence induced changes in oxytocin expression in male and female rats. Oxytocin decreased sIPSC amplitudes in both naïve and dependent male animals, but blocked alcohol‐induced enhancement of sIPSC frequency in dependents only, indicating both pre‐ and post‐synaptic effects of oxytocin on GABAergic transmission. Pretreatment with an oxytocin antagonist blocked the effects of oxytocin and restored the alcohol enhancement in sIPSC frequency. In female naïve rats, oxytocin has no effects on sIPSCs alone, and does not block the alcohol enhancement of sIPSC frequency, but in alcohol dependent females, oxytocin increases sIPSC frequency alone and additional alcohol increases sIPSC frequency even further. These results provide important insight into the role of oxytocin in CeA neuronal functioning and the sexually dimorphic changes in this system that contributes to the transition to alcohol dependence. This research highlights the potential role of oxytocin in the development of novel targeted therapeutics for the effective treatment of AUDs.

Support or Funding Information

NIH/NIAAA F32: AA025262, T32: AA007456, R01: AA017447, AA006420, AA015566

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of the discriminative stimulus effects of the delta‐opioid receptor agonist, SNC80, in rats
Authors: Matthews Ashleigh L. , Rice Kenner C. , Jutkiewicz Emily M.
Published: June 2021

Delta opioid receptor (DOR) agonists produce antihyperalgesia and antidepressant‐like effects, but they also induce convulsions limiting their clinical development. Recently, some work has shown that not all DOR agonists produce convulsions in rodents. Therefore, the current study aimed to further evaluate the pharmacological properties of these DOR ligands using a drug discrimination paradigm in male Sprague‐Dawley rats. The drug discrimination assay is useful for identifying compounds with similar interoceptive properties and evaluating the pharmacological activity of ligands in vivo. For this study, rats were trained to discriminate 3.2 mg/kg SNC80 from saline on a fixed ratio (FR) 10 schedule of reinforcement in a food‐reinforced, drug discrimination paradigm. Substitution patterns with purported DOR agonists AZD2327, DPI287, KNT127, and MMP2200 were evaluated as well as profiles of antagonism following pretreatment with naltrindole. Agonists that are structurally similar to SNC80, AZD2327 and DPI287, generalized to SNC80 while structurally unrelated agonists, KNT127 and MMP2200, did not. Morphine, ketamine, desipramine, cocaine, and nicotine failed to generalize to SNC80 up to doses that suppressed responding; however, amphetamine produced partial generalization to the discriminative stimulus effects of SNC80. The DOR antagonist naltrindole produced parallel, rightward shifts in the SNC80 dose effect curve. Overall, these data suggest that interoceptive effects differ among DOR agonists, which may contribute to their distinct profiles of behavioral activity and potentially diverse mechanisms of action. Future studies will further probe the pharmacological and/or signaling mechanisms that differentiate the behavioral effects of DOR agonists.

Support or Funding Information

This work is supported in part by R01 DA042092 (EMJ) and the IRPs of NIDA and NIAAA (KCR).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Leukemia Inhibitory Factor Modulates the Post‐Stroke Immune Response in Rats
Authors: Davis Stephanie M. , Collier Lisa A. , Leonardo Christopher C. , Ajmo Craig T. , Kopper Timothy J. , Gensel John C. , Pennypacker Keith R.
Published: June 2021

Objective

To determine if leukemia inhibitory factor (LIF) administration after permanent focal stroke decreases post‐stroke neuroinflammation.

Methods

Focal ischemia was induced in 3 month old male and 18 month old male/female Sprague‐Dawley using the middle cerebral artery occlusion (MCAO) procedure. Animals were treated with PBS or LIF at 6, 24, and 48 h after MCAO (125 μg/kg). Animals were euthanized at at 72 h post‐MCAO. Protein levels of CD11b, IL‐12 p40, IFNγ, and IP‐10 were measured in splenic tissue using immunoblotting and/or ELISA. Isolectin staining and immunoblotting were used to detect levels of CD11b+ macrophages/microglia in the brain. Murine bone marrow derived macrophages (BMDMs) stimulated with LPS (50 ng/ml) and IFNγ (20 ng/ml) in the presence or absense of LIF and IL‐12 p40/IL‐10 levels were detected in the supernatant.

Results

LIF treatment significantly increased the average spleen weight at 72 h after MCAO compared to PBS‐treated rats (p<0.05) and decreased splenic LIFR expression compared to sham operated (p<0.01) and PBS‐treated (p = 0.0682) rats. LIF‐treated rats showed significantly decreased levels of IL‐12 p40 (p <0.05) and IFNγ (p = 0.0644) in splenic tissue at 72 h after MCAO compared PBS‐treated animals. LIF treatment also counteracted the significant increase in splenic IP‐10 expression observed in the PBS‐treated rats (*p<0.01 compared to sham‐operated rats). CD11b levels were significantly lower in the spleens of LIF‐treated (p<0.001) and PBS‐treated (p<0.01) rats compared to sham rats. However, LIF treated significantly decreased normalized CD11b protein levels (p<0.05) and decreased isolectin reactivity in the brain at 72 h post‐MCAO compared to PBS‐treated animals. BMDMs treated with LIF prior to IFNγ and LPS stimulation released significantly lower levels of IL‐12 p40 and significantly higher levels of IL‐10 (p ≤ 0.05) compared to BMDMs treated with PBS prior to LPS+IFNγ stimulation.

Conclusion

LIF confers anti‐inflammatory signaling after stroke through modulation of the IL‐12 p40/IFNγ/IP‐10 axis in splenocytes and decreasing the infiltration of CD11b+ monocytes/macrophages into the ischemic brain.

Support or Funding Information

Project Number: 5R01NS091146‐03 (NINDS; PI: Pennypacker)Project Number: 55R01NS091582‐03 (NINDS; PI: Gensel)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Schematic diagram of the neural protective role of IMMH004 after TGCI/R Pharmacological Induction of Mitochondrial Biogenesis using the β2‐Adrenoreceptor Agonist Formoterol for the Treatment of Spinal Cord Injury
Authors: Scholpa Natalie E. , Wang Wenxue , Corum Daniel , Narang Aarti , Tomlinson Stephen , Schnellmann Rick G.
Published: June 2021

A hallmark of the progressive cascade of damage referred to as secondary spinal cord injury (SCI) is vascular disruption resulting in ischemia and decreased oxygen delivery, which directly reduces the ability of mitochondria to maintain homeostasis. This leads to loss of ATP‐dependent cellular functions, Ca2+ overload, excitotoxicity and oxidative stress, further exacerbating injury. While therapeutics targeting restoration of single facets of mitochondrial function have proven largely ineffective post‐SCI, comprehensively addressing mitochondrial dysfunction via pharmacological stimulation of mitochondrial biogenesis (MB) is an underexplored strategy. The FDA‐approved, highly selective and potent β2‐adrenoreceptor (ADRB2) agonist formoterol is an effective inducer of MB in multiple organ systems in mice, including the spinal cord, as evidenced by increased mitochondrial DNA content and PGC‐1α expression. This study examined the effects of formoterol‐induced MB on recovery from SCI using a force‐controlled pneumatic impactor‐induced contusion model. Female C57bl/6 mice were subjected to moderate SCI (80 Kdyn) followed by daily formoterol administration (0.1 mg/kg, i.p) beginning 1 or 8 h post‐injury and continuing until euthanasia. SCI resulted in decreased mitochondrial protein expression, including PGC‐1α, in the injury and peri‐injury sites as early as 3 d after injury. Formoterol treatment attenuated this early decrease in PGC‐1α and restored downstream mitochondrial proteins to levels similar to that of sham controls by 15 d post‐SCI. Formoterol‐treated mice also exhibited less histological damage than vehicle‐treated mice 3 d after injury, namely reduced loss of white matter and structural preservation of the central canal and anterior median fissure. Importantly, as measured by the Basso‐Mouse Scale, the locomotor ability of formoterol‐treated injured mice reached twice that of vehicle‐treated SCI mice by 15 d (4.5 v 2), depicting nearly 50% recovery. Interestingly, a comparable degree of functional recovery was observed when formoterol treatment initiation was delayed until 8 h post‐SCI. These data provide evidence of ADRB2‐mediated MB as a therapeutic avenue for the treatment of SCI.

Support or Funding Information

National Institutes of Health National Institute of General Medical Sciences, Biomedical Laboratory Research and Development Program of the Department of Veterans Affairs, South Carolina Spinal Cord Injury Research Fund

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sodium butyrate exerts neuroprotective effects in spinal cord injury.
Authors: Lanza Marika , Campolo Michela , Casili Giovanna , Filippone Alessia , Cuzzocrea Salvatore , Esposito Emanuela
Published: June 2021

Sodium butyrate (SB) is a dietary microbial fermentation product of dietary fiber and serves as an important neuromodulator in the central nervous system. Recent experimental evidence has suggested potential therapeutic applications for butyrate, including its utility in treating metabolic and inflammatory diseases. The aim of the present study was to evaluate the potential beneficial effects of SB in a mouse model of spinal cord injury (SCI) and its possible mechanism of action. SCI was produced by extradural compression for 1 min of the spinal cord at the T6–7 level using an aneurysm clip, and SB (10–30–100mg/kg) were administered by oral gavage 1 and 6 h after SCI. For locomotor activity, study mice were treated with SB once daily for 10 days. Morphological examination was performed by light microscopy through hematoxylin‐eosin (H&E) staining. In addition, NF‐kB, IkB‐alfa, COX2 and iNOS expression were assayed by western blot analysis and IL‐1beta and TNF‐alfa levels by immunohistochemistry analysis. The results showed that SB treatment significantly ameliorated histopathology changes and improved recovery of motor function changes in spinal cord injury in dose‐dependet manner. Moreover, from the results obtained, SB modulated NF‐kB pathway showing a reduction in cytokines expression. This study showed that SB exerts neuroprotective effects on spinal cord injury and anti‐inflammatory properties. The observed neuroprotective action suggests that SB may serve as a potential candidate for future treatment of spinal cord injury.

Support or Funding Information

No founding

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 Cannabis use on the Antipsychotic Therapy of Schizophrenia Symptoms
Authors: Thomas‐Brown Peta‐Gaye Latoya , Martin Jacqueline Simone , Sewell Clayton A. , Abel Wendel D. , Gossell‐Williams Maxine D.
Published: June 2021

Objectives

To assess the impact of Cannabis usage on risperidone and haloperidol therapy of schizophrenia symptoms.

Methods

Male subjects, ≥18 years, admitted to the University Hospital of the West Indies psychiatric ward between February‐December 2013, and diagnosed with schizophrenia were recruited for the study. Consented subjects were prescribed either risperidone‐ or haloperidol‐monotherapy for 7‐days and oral doses converted to chlorpromazine equivalents (CPZE). Subjects were classified as Cannabis users (CU) or non‐users (non‐CU), with presence/absence of Cannabis metabolite in urine samples. Blood samples were collected on days 1&7 for assessment of cannabinoid and antipsychotic concentrations, respectively, using ultra performance liquid chromatography tandem mass spectrometry (UPLC/MS/MS). Psychotic symptoms were assessed with the Brief Psychiatric Rating Scale (BPRS) on days 1&7 and changes in the scores used as a measure of efficacy of the antipsychotic therapy. Inferential statistical analysis involved non‐parametric tests, expressed as median and IQR, with p<0.05 considered significant.

Results

Forty‐Six Afro‐Caribbean males were assessed, with median (IQR) age of 25 (8) years. Majority (n=33; 72%) were CU, with Cannabis‐positive urinalysis, confirmed with the quantification of Δ9‐tetrahydrocannabinol (THC) at 3.0 (8.0) ng/mL and Carboxylated‐THC at 51.0 (53.0) ng/mL in serum samples. CU presented with more severe BPRS scores [61 (33) vs 33 (19); p<0.000] than non‐users. Additionally, more CU displayed depression (60%; r=0.403, p=0.001) and motor retardation (79%; r=0.595, p=0.002) which were positively correlated to serum THC concentration. After seven days of antipsychotic therapy, both CU and non‐CU showed significant clinical improvement of schizophrenia symptoms (≥20% reduction in BPRS scores) with risperidone or haloperidol at CPZE doses ≥300 mg/day. A significantly higher oral CPZE dose of haloperidol was prescribed to CU than non‐CU [1000 (250) vs 500 (500) mg/day; p=0.035]. However, serum concentration of haloperidol was significantly less for CU than non‐CU [3 (2) vs 8 (3) ng/mL; p=0.044]. In contrast, oral doses [300 (75) vs 300 (0); p=0.554] and serum concentration of risperidone [12 (5) vs 13 (4) ng/mL; p=0.739] were equivalent between the groups.

Conclusion

Cannabis users display more severe BPRS‐assessed schizophrenia symptoms compared to non‐users, which can be significantly improved after 7 days of antipsychotic therapy. However, Cannabis use reduces the potency of haloperidol, thus, a significantly higher oral dose is required to adequately treat schizophrenia symptoms. Further studies are recommended.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glycogen Synthase Kinase‐3β Modulation of Basal Aryl Hydrocarbon Receptor Protein Levels in HeLa Cells
Authors: Yang Yujie , Chan William K.
Published: June 2021

The aryl hydrocarbon receptor (AHR) is a transcription factor involved in many cellular processes. It has been well accepted that it plays important roles in development, metabolism, immune response, and carcinogenesis. Presently the only known regulation of AHR protein is mediated through proteasomal degradation after ligand activation. In this study, we investigate the mechanisms that govern the AHR protein levels without ligand treatment. The ligand‐free AHR resides in the cytosol and is accompanied by XAP2, Hsp90, and its co‐chaperone p23. We previously discovered that knockdown of p23 can lower the AHR protein levels as well as its message levels. Close examination of the AHR sequence unveils a putative signal for glycogen synthase kinase‐3β (GSK‐3β) phosphorylation which may lead to protein degradation. Treatment of GSK‐3β inhibitors tideglusib and lithium chloride increases the AHR protein levels up to about 2‐fold in wild type HeLa cells but not in p23 knockdown stable HeLa cells. These results suggest that the AHR protein levels can be regulated by GSK‐3β only in the presence of the wild type content of p23. Downregulation of the AHR protein levels by GSK‐3β and p23 appears to be mediated through distinct mechanisms. We will address whether this GSK‐3β effect is p23‐dependent. Additional mechanistic data will be presented to explain the underlying mechanisms.

Support or Funding Information

NIEHS, R15ES023104

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human Antigen R (HuR) Regulates Structure and Function of Brown Adipose Tissue
Authors: Lanzillotta Lindsey , Anthony Sarah R. , Yamamoto Kaila , Jones Shannon , Taylor Zachary , Yoshida George , Owens A. Phillip , Tranter Michael
Published: June 2021

Human antigen R (HuR) is an RNA binding protein widely expressed throughout the body, including in both white (WAT) and brown (BAT) adipose tissue. Recent work from our lab has shown that adipocyte‐specific HuR knockout (adipo‐HuR−/−) mice gain less weight following a high fat diet (HFD) compared to wild type (WT) controls. In this study, we focused on the role of HuR deletion on the function of BAT, which mediates non‐shivering thermogenesis.

Histological analysis revealed that, while the BAT of the chow fed adipo‐HuR−/− mice appeared normal, the BAT of the HFD fed adipo‐HuR−/− mice appears less dense with a cell size that more closely resembles WAT. Upon further examination of BAT function, our results show that, when subjected to cold stress (4°C), adipo‐HuR−/− mice are less cold tolerant compared to their WT counterparts. We also show that, compared to the WT mice, adipo‐HuR−/− mice have decreased mitochondrial density and expression of uncoupling protein‐1 (UCP‐1) in their BAT. This may be caused by a HuR‐dependent decrease in expression of peroxisome proliferator‐activated receptor gamma coactivator 1 alpha (PGC‐1a), which plays a key role in mitochondrial biogenesis and is upregulated during exposure to cold (4°C).

In conclusion, our results indicate that HuR regulates both the structure and function of BAT, at least in part through modulation of PGC‐1a mRNA expression.

Support or Funding Information

This work was supported by a University of Cincinnati Heart, Lung, and Vascular Institute Near Horizons Grant (MT, APO).

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