bluuebunny/crossref_metadata_2025_split huggingface dataset:
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.METHODSWe 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.RESULTSWe 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 InformationSupported 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 InformationThis 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 InformationThis 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 InformationThis 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 InformationThis 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 InformationNIH/NIAAA F32: AA025262, T32: AA007456, R01: AA017447, AA006420, AA015566This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.