bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Ketogenic Diet on p70S6k Phosphorylation in Young Adult and Old Soleus Muscle
Authors: Bennett Shantelle L.H. , Smith Andrew H. , Ramirez J. Lucas , Thomson David M. 
Published: June 2021

A ketogenic diet (KD) has been proposed as a treatment for many conditions including obesity, diabetes, epilepsy, and Parkinson's Disease. Various studies have shown its effects on health and lifespan in model organisms suggesting that it is beneficial for many individuals. Some concern exists that KD might negatively impact anabolic signaling through the mTOR pathway, but the molecular signaling effects of the diet remain largely unexplored, particularly in skeletal muscle. The purpose of this study, therefore, was to determine how KD affects downstream mTOR signaling in skeletal muscle from young and old rats. Young adult (YA; 5 mo.) and old (O; 28 mo.) male Fisher 344 rats were isocalorically fed either a ketogenic diet (KD; Envigo Teklad Custom Diet, TD.10911; 22.4% protein, 77.1% fat, 0.5% carbohydrates) or standard diet (STD; Envigo Teklad Rodent Diet, 8604; 32% protein, 14% fat, 54% carbohydrate) for four weeks. Soleus muscles were harvested and assessed for muscle size and effects on p70 ribosomal protein S6 kinase (S6k) phosphorylation. Soleus muscles from YA KD rats were ~11% smaller than for corresponding YA STD rats, and O KD soleus muscles were ~6% larger vs. O STD muscles, though neither of these differences were significant. Phosphorylation of S6k was significantly (p < 0.05) decreased 75% in YA KD vs. YA STD. S6k phosphorylation was reduced 88% in O muscles, but, in contrast to the YA muscles, was increased by ~400% by KD vs. STD in O muscles. Based on this evidence, we conclude that in slow‐twitch soleus muscles, administration of KD can decrease skeletal muscle mTOR signaling in YA muscles and increase its signaling in O muscles.Support or Funding InformationThis work was supported by a Brigham Young University Gerontology Program award.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evidence of endothelial‐to‐mesenchymal transition in an animal model of pulmonary arterial hypertension
Authors: Morrow K. Adam , Brown Jonathan R. , Morrow Rebekah L. , Snyder Emily A. , Vasauskas Audrey A 
Published: June 2021

Pulmonary arterial hypertension (PAH), a disease of the small pulmonary arteries, is characterized by pulmonary vasoconstriction, vascular cell proliferation, and vascular remodeling leading to right ventricular hypertrophy and failure. The remodeled vessels exhibit signs of endothelial layer damage and smooth muscle cell proliferation. It is also well‐established that mutations in the bone morphogenetic protein receptor 2 (BMPR2) predispose individuals to development of PAH. Recent studies in PAH mouse models and in cultured human pulmonary artery endothelial cells (where BMPR2 has been experimentally knocked down) suggest that endothelial cells involved in the vascular remodeling of PAH undergo transdifferentiation. Here, we show that pulmonary artery endothelial cells (PAECs) isolated from the rat Sugen (SU516)/hypoxia model of pulmonary arterial hypertension exhibit phenotypic changes characteristic of endothelial‐to‐mesenchymal transition (EndMT). Preliminary data also suggest decreased BMPR2 expression in the PAH model PAECs. Cells were obtained from the University of South Alabama Center for Lung Biology (CLB). Briefly, Fisher 344 rats were injected with SU5416 (20mg/kg) or vehicle control and exposed to hypoxia (10% O2 or normoxia for control animals) for three weeks and returned to normoxia for at least two weeks. PAECs were harvested using established CLB protocol. PAECs from the PAH animals (SURAT) exhibited a significant increase in alpha smooth muscle actin (αSMA) expression as compared to the control PAECs, as evidenced by immunocytochemistry and real time PCR. These cells retained expression of endothelial cell markers von Willebrand Factor (vWF) and platelet endothelial cell adhesion molecule (PECAM‐1). However, expression of both PECAM‐1 and vWF was decreased in the SURAT cells compared to control PAECs. Expression of tight junctional protein‐1, vascular adhesion molecule‐1 (VCAM‐1), intracellular adhesion molecule‐1 (ICAM‐1), claudin‐1, and selectin was also decreased in the SURAT PAECs as compared to control cells. These data suggest that PAECS exhibit signs of cellular reprogramming in Sugen/hypoxia‐treated animals, and also provide insight into a cellular model that directly correlates with a well‐established animal model of PAH.Support or Funding InformationSupported by NIH1R15HL137135‐01A1This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
LDLR‐Related Protein 1 Increases Cytokine Sensitivity – Implications for Recovery after Brain Damage
Authors: Ahmad Sadiya Noor , Reed Pam , Sprague Shane , Sayre Naomi 
Published: June 2021

The limited treatment options for ischemic stroke patients have resulted in stroke being a leading cause of death and the primary cause of long‐term disability in the U.S. Finding effective treatment options requires a better fundamental understanding of the ongoing processes that contribute to poor long‐term outcome.Expression of Apolipoprotein E4 predisposes stroke patients to poor long‐term outcome. This study aims to test one possible mechanism by which ApoE4 contributes to cognitive decline after stroke. Here, we examine the effect of a major ApoE4 receptor, low‐density lipoprotein receptor related protein 1 (LRP1) on sensitivity to stress in astrocytes.LRP1 binds and moves extracellular ligands and plasma membrane proteins into the endocytic system. Others have shown that LRP1 regulates cell‐surface TNF receptor (TNFR1) in non‐astrocytic cells. We propose That LRP1 similarly regulates TNFR1 in the central nervous system to attenuate inflammatory response after stroke. Studies have shown that ApoE4 slows the recycling of endocytic LDL receptors. We hypothesize that ApoE4 inhibits the ability of LRP1 to remove TNFR1 from the plasma membrane. This is expected to increase cytokine sensitivity, resulting in worse outcome after stroke.We investigated the effect of LRP1 on astrocyte TNFα signaling and response in immortalized ApoE null mouse astrocytes subjected to lentiviral‐mediated knockdown of LRP1. The astrocyte response to TNFα stimulation was tested in a time dependent manner using Western blotting of NFkB pathway components, which are the downstream mediators of TNFα signaling. We also tested astrocyte viability after prolonged TNFα stimulation using Alamar Blue reagent. We found that LRP1 deficient cells have increased phosphorylation of NFkB upon TNFα stimulation, and that loss of LRP1 resulted in significant loss of astrocyte viability after prolonged stimulation. Altogether, our results indicate that loss of LRP1 renders astrocytes more sensitive to TNFα. Future experiments will focus on testing the influence of LRP1 on recovery after middle cerebral artery occlusion in mice.These studies will elucidate how astrocyte‐LRP1 contributes to outcome after stroke, and helps us to understand one potential way that ApoE4 exerts pathological effects. A better understanding of the long‐term processes after stroke will allow identification of therapies which improve the morbidity and mortality associated with stroke.Support or Funding InformationInstitutional National Research Service Award, T32 NIH/NCATS, TL1TR002647, funded by National Institutes of Health/National Center for Advancing Translational SciencesThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Tunicamycin‐induced ER Stress Effect on Cardiac Contractility
Authors: Alabdaljabar Mohamad Saleh , Valencia Sara Osorio , Han Young Soo , Sieck Gary C. 
Published: June 2021

In cardiomyocytes, the endoplasmic reticulum (ER) plays a dynamic role in excitation‐contraction coupling by regulating cytosolic Ca2+ concentration ([Ca2+]cyt). Under pathological conditions the ER accumulates unfolded proteins triggering a homeostatic ER stress response to minimize and limit protein unfolding (via chaperone proteins) and accumulation (by decreasing synthesis and increasing degradation). The interactions between ER stress, [Ca2+]cyt regulation and downstream contractile responses remain unknown, although such interactions may underlie the etiology of various cardiovascular diseases. We hypothesized that in cardiomyocytes ER stress alters the regulation of [Ca2+]cyt and reduces cardiac contractility by decreasing myofilament Ca2+ sensitivity. In isolated rat cardiomyocytes, ER stress was induced by tunicamycin, a naturally occurring antibiotic that induces ER stress by inhibiting N‐linked glycosylation. In one set of experiments, we used Western blotting techniques to evaluate the effects of ER stress on phosphorylation of cTnI at Ser23/24. In a second set of experiments, we evaluated the impact of tunicamycin on excitation‐contraction coupling by simultaneously measuring [Ca2+]cyt (using Fura‐2 AM ratiometric imaging) and contractile (sarcomere length (SL) shortening) responses to electrical field stimulation using an IonOptix system. Tunicamycin induced an increase in phosphorylation of cTnI by 3–6 h. The evoked [Ca2+]cyt transient was relatively unaffected by tunicamycin while the contractile response was markedly reduced. This resulted in a rightward shift in the phase loop relationship between [Ca2+]cyt and SL shortening indicating reduced myofilament Ca2+ sensitivity. We conclude that ER stress may underlie contractile dysfunction in a variety of cardiac diseases.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Induction of IL‐10 in Bone‐Marrow Derived Macrophages by Helicobacter Hepaticus Supernatant is Mediated by Gpr109a
Authors: Al‐odaini Asma , Singh Nagendra 
Published: June 2021

Interleukin 10 (IL‐10) is an anti‐inflammatory cytokine, and its polymorphism is linked to inflammatory bowel diseases (IBDs) in humans. Interactions between gut microbiota and the host's the immune cells play a key role in shaping immune responses and intestinal health. Helicobacter hepaticus is a member of mouse gut microbiota that is tolerated by the host but can induce colonic inflammation in mice in absence of an intact IL‐10 signaling. H. hepaticus secretes a polysaccharide that signals through Toll‐like receptor 2 (TLR2) to induce IL‐10 production. Gpr109a is a G‐protein coupled receptor that is expressed at a very high level on innate immune cells, and it is thought to interact with gut microbe‐derived molecules to induce production of anti‐inflammatory cytokines and suppress colonic inflammation. It is hypothesized that H. hepaticus induction of IL‐10 through TLR2 is mediated by Gpr109a. To test whether Gpr109a is involved in IL‐10 induction by H. hepaticus secretions, bone‐marrow derived macrophages (BMDMs) of WT and Gpr109a KO mice were stimulated by H. hepaticus supernatant then IL‐10 mRNA and protein levels were determined. WT BMDMs were found to produce significantly more IL‐10 mRNA and protein compared to Gpr109a KO BMDMs. In addition, ERK1/2 is a protein complex that gets activated when the TLR2 is stimulated by H. hepaticus to induce IL‐10. We tested whether ERK1/2 activation by H. hepaticus supernatant is dependent on Gpr109a by using an immunoprecipitation assay for total and pERK1/2 upon stimulation with H. hepaticus supernatant for WT and Gpr109a KO BMDMs. Our data suggests that Gpr109a plays a critical role in activation of ERK1/2 downstream of TLR2 signaling leading to induction of IL‐10. These findings will be of a value in delineating the molecular mechanism of Gpr109a signaling and investigating Gpr109a role in preventing H. hepaticus induced colitis and intestinal cancers.Support or Funding InformationThis projected was funded by Augusta University (The STAR Program) and performed in Dr. Nagendra Singh's laboratory and under his supervision with technical training and support by Brinda Bhatt and Dr. Huabin Zhu.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aerobic Training Reduces Immune Cell Recruitment and Cytokine Levels in Adipose Tissue in Obese Mice
Authors: Silva Albená Nunes , Lacerda Debora Romualdo , Moraes Michele Macedo , Costa Kátia Anunciação , Rodrigues Débora Fernandes , Sabino Josiana Lopes , de Souza Cordeiro Letícia Maria , Pinho Vanessa , Teixeira Mauro Martins , Wanner Samuel Penna , Soares Danusa Dias , Ferreira Adaliene Versiani Matos 
Published: June 2021

Obesity is associated with an energy imbalance that results from excessive energy intake, low diet quality and a sedentary lifestyle. In this regard, the increased consumption of a high‐refined carbohydrate diet (HC) is strongly related to higher adiposity and low‐grade inflammation. Aerobic training is a well‐known non‐pharmacological intervention to treat obesity and metabolic disturbances. However, the mechanisms through which aerobic training ameliorates the low‐grade inflammation induced by the HC diet need to be further investigated. Herein, our hypothesis was that aerobic training would decrease the recruitment of leukocytes in the adipose tissue thereby reducing the levels of cytokines and improving metabolism in mice fed the HC diet. Male Balb/c were assigned to the following groups: control non‐trained (C‐NT), control trained (C‐T), HC‐NT and HC‐T. Mice were submitted to moderate‐intensity training sessions that consisted of running 60 min/day for 8 weeks. The intravital microscopy technique was performed in vivo in anesthetized mice to visualize the microvasculature of the adipose tissue. The HC diet induced obesity and increased the influx of immune cells into the adipose tissue. In contrast, HC‐T mice presented a lower adiposity and adipocyte area. Furthermore, HC‐T mice showed an increased resting energy expenditure, a decreased recruitment of immune cells in the adipose tissue, reduced cytokine levels, and ameliorated hyperglycemia and fatty liver deposition relative to HC‐NT mice. Collectively, our data enhance the understanding about the anti‐inflammatory effect of aerobic training and shed light on the adipose tissue‐mediated mechanisms by which training promotes a healthier metabolic profile.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Resveratrol protects UVB‐irradiated keratinocytes by upregulating HO‐1 expression and improving mitochondrial function via endogenous CO signaling
Authors: Averilla Janice Nullan , Oh Jisun , Kim Jong‐Sang 
Published: June 2021

Excessive exposure to ultraviolet (UV) radiation has been reported to promote uncontrolled reactive oxygen species (ROS) production, particularly in the mitochondria, and thus implicated in the prevalence of skin injuries. Considering the antioxidative effect of resveratrol (RES) in reducing UV‐induced ROS production in human keratinocytes (HaCaT cells) and the role of carbon monoxide (CO) in regulating oxidative stress, we hypothesized that RES may promote cellular antioxidant response by activating heme oxygenase‐1 (HO‐1) which can generate its enzymatic product, CO, in UVB‐irradiated keratinocytes. To test our hypothesis, we treated HaCaT cells with CO‐releasing molecule 2 (CORM‐2) (source of CO) and RES either independently or in combination and irradiated the cells with UVB. Co‐treatment of CO with RES upregulated HO‐1 expression and subsequently reduced intracellular ROS in response to UV irradiation. Moreover, UVB treatment alone induced mitochondrial biogenesis, but treatment with CO and RES did not increase it further. With UVB treatment, RES showed enhanced mitochondrial respiration while CO did not instigate any effect; however, independent or co‐treatment mitigated mitochondrial ROS production. Taken together, our data support the impression that endogenous CO generated from RES‐mediated activation of HO‐1 may play a crucial role in cellular antioxidant response and mitochondrial function.Support or Funding InformationThis work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry, and Fisheries (IPET) through the High Value‐Added Food Technology Development Program, funded by the Ministry of Agriculture, Food, and Rural Affairs (Grant No. 116026‐03‐2‐HD020), Republic of Korea.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Effects of a Bout of Moderate‐intensity Physical Exercise on Adipose Tissue Inflammation in Lean and in Obese Mice
Authors: Silva Albená Nunes , Lacerda Debora Romualdo , Rodrigues Débora Fernandes , Silveira Ana Leticia Malheiros , Moraes Michele Macedo , Pinho Vanessa R , Teixeira Mauro Martins , Wanner Samuel Penna , Soares Danusa Dias , Ferreira Adaliene Versiani 
Published: June 2021

Inflammation is an adaptive response that is triggered by the noxious stimuli in order to maintain the homeostasis. Previous studies have shown that higher lipolysis rate after fasting is associated with the influx of leukocytes and increased cytokine levels in the epididymal adipose tissue (EAT). Fat mobilization is also highly activated during and post‐exercise. In spite of substantial evidences having supported the link between inflammation and exercise, the association between EAT, cytokines and acute exercise is less explored. Herein, we characterized the EAT inflammation following exercise in lean and obese mice. We used in vivo confocal imaging from EAT vessel to display the higher recruitment of neutrophils 0 and 6 hours post‐exercise in lean mice. Additionally, we found that exercise enhanced the levels of IL‐6 and TNF‐alpha in the EAT which were correlated with increased serum free fatty acids in lean mice. Interestingly, obese mice subjected to exercise showed lower pro‐inflammatory level of cytokines in the EAT and lost the correlation between IL‐6, TNF‐alpha and free fatty acids. Furthermore, obese mice increased IL‐10 24h post‐exercise. Together, our data show that exercise induces inflammation in the EAT in lean mice. These responses may have a physiological purpose and be important to stimulate fat breakdown. Conversely, exercise in obese mice promoted a reduction in pro‐inflammatory mediators and elevated IL‐10. Despite of the low‐grade inflammation being associated with metabolic dysfunctions, at least in part, the inflammatory milieu in EAT may be also important to regulate fat remodeling induced by exercise.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
IIntracellular BiP/GRP78 Mediates Endothelial Cell Permeability and Inflammation Associated with Acute Lung Injury
Authors: Leonard Antony , Su Pei‐Yi , Millar Michelle , Dubey Nikhil , Yule David , Rahman Arshad , Fazal Fabeha 
Published: June 2021

The role of endoplasmic reticulum (ER) chaperone and signaling regulator BiP/GRP78 in acute inflammatory injury, particularly in the context of lung endothelium, is poorly defined. Recent studies have shown that BiP/GRP78 not only resides in the ER but is present in extra‐ER regions as well, and perform different functions. In this study, we monitored the effect of intracellular versus cell surface BiP/GRP78 in EC permeability and inflammation associated with acute lung injury (ALI). Studies were performed using SubAB, a holoenzyme that cleaves and specifically inactivates intracellular BiP/GRP78 versus SubA, which cleaves and inactivates only the cell surface BiP/GRP78. Stimulation of endothelial cell (EC) with proinflammatory mediator thrombin showed a characteristic time‐dependent change in TER (trans‐endothelial resistance), a real time measurement of EC permeability, which was protected in the presence of SubAB but not in the presence of SubA and its inactive mutant SubAA272B. In order to further dissect the mechanism of thrombin‐mediated EC permeability, we determined the role of BiP/GRP78 in mediating Ca2+ signaling, a critical determinant of EC permeability. Interestingly, SubAB significantly reduced thrombin‐induced Ca2+ release from the ER stores as well as Ca2+ entry from the extracellular medium; however SubA and SubAA272B failed to mediate the above response. Furthermore, mechanistic analysis revealed that thrombin‐induced activation of STIM‐1 (Stromal Interaction Molecule 1) via Y361 phosphorylation, which stimulates Ca2+ entry across the plasma membrane, was blocked in the presence of SubAB. In addition, thrombin‐induced phosphorylation of eNOS at serine 1177 as well as the association of phosphorylated eNOS with p190GAP, which is a major mechanism responsible for AJ disassembly resulting in increased EC permeability, was significantly blocked by SubAB. Interestingly, thrombin‐induced inflammatory responses were not affected in the presence of SubA as opposed to SubAB. Together, these data highlight a novel role of intracellular BiP/GRP78 in mediating EC barrier disruption and inflammation associated with ALISupport or Funding InformationNIH (HL096907 and HL130870)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Co‐localization of Isolectin B4 (IB4), Ionized Calcium‐Binding Adapter Molecule 1 (Iba1), and Von Willebrand Factor (vWF) Immunostaining in the Mid‐Cervical Spinal Cord After Spinal Injury
Authors: Fusco Anna F , Sunshine Michael D , Streeter Kristi A , Wollman Lila B , Jorgensen Marda L , Fuller David D 
Published: June 2021

IB4 immunostaining is often used to histologically identify thin fiber non‐peptidergic afferent neurons in the spinal cord. However, IB4 can also identify both endothelial cells and microglia. In studies of adult Sprague Dawley rats with chronic (12 weeks) cervical (C2) hemisection injury (Hx; N=10), we noted robust IB4 immunostaining in mid‐cervical dorsal root ganglia cell bodies as well as spinal cord lamina II, indicating labeling of thin fiber afferents. We also observed distinct clusters of IB4+ cells in the mid‐cervical spinal cord ipsilateral to Hx in the area of the corticospinal tract (N=6 of 10 rats), and in ventrolateral (N=9 of 10) and ventromedial white matter (N=9 of 10). Incubation of tissues with primary antibodies for both IB4 and Iba1 (which can label activated microglia) revealed clear co‐localization of staining in these regions. Spinal intact rats (N=5) had robust IB4 staining in lamina II, but staining was not observed in the corticospinal tract, or ventrolateral or ventromedial white matter. The IB4‐Iba1 co‐localization after Hx likely indicates neuroinflammation and degeneration of damaged tracts. We also observed that IB4 immunostaining broadly dispersed throughout white and gray matter, in a pattern consistent with blood vessel endothelium labeling. To verify this, tissues were co‐incubated with primary antibodies against IB4 and vWF, a marker of endothelial cells. Co‐localization of vWF and IB4 immunostaining was detected throughout the mid‐cervical cord, and IB4 staining intensity was decreased in ventral gray matter caudal to Hx (P<0.05 vs. spinal intact). Reduction of IB4 intensity is consistent with vascular damage and regional tissue hypoxia. We conclude that following chronic C2Hx in adult rats, immunostaining with IB4 can identify blood vessel endothelial cells and activated microglia.Support or Funding Information1R01HL139708‐01A1 (DDF), OT2 OD023854, T32‐HD043730 (MDS), K99 HL143207‐01 (KS).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.