bluuebunny/crossref_metadata_2025_split huggingface dataset:
Amelioration of N‐acetyl‐1,4‐benzoquinone imine‐Induced Hepatotoxicity in Rats by Aqueous Leaf Extract of Annona Muricata (Soursop)
Authors: Nosiri Chidi Ijeoma , Anyanwu Chukwuma , Okwara Elizabeth Nnenna 
Published: June 2021

The leaf of A.muricata is traditionally used to boost the liver and kidney. Acetaminophen (ACT), an analgesic and over the counter drug is usually abused. Overdose of it could lead to hepatic oxidative stress where the intermediate N‐acetyl‐1,4‐benzoquinoneimine which is rapidly detoxified by Glutathione S‐transferase becomes too much for this enzyme to handle. This mechanism is important in the development of acetaminophen toxicity. 20 albino rats randomly divided into 4 groups of 5 rats were used for the study. (Group 1:normal saline, group 2: ACT alone (400 mg/kg p.o), group 3: ACT and plant extract (400mg/kg p.o) and group 4: ACT and extract 800 mg/kg p.o. For the results of ALT (Alanine amino transferase) and AST (Aspartate amino transferase) levels, group 2 showed a significant increase compared to the control and treated groups. ALT for the control group was 17.00±1.00, group 2 value: 35.00±1.00, group 3 value: 31.00±1.00 and group 4 values: 21.00±1. For AST, the control value gave 26.00±1.00, group 2 value: 45.00±2.50, group 3 value: 38.00±1.00 and group 4 value: 29.00±1. For ALP (Alkaline phosphatase), the control value: 65.16±1.44, group 2 value: 70.05±0.81, the group 3 value 67.79±0.58 while group 4: 64.37±2. In all, the treated groups dose dependently ameliorated the hepatotoxicity shown by the levels of the liver enzymes. The high dose of (NAPQI) may have reacted with hepatic glutathione (GSH) leading to low concentration of total hepatic GSH that generated the production of free radicals which destroyed the liver cells causing the liver enzymes to leak into the blood. The histopathological effect also showed moderate to severe portal inflammation in group 2 while groups 1,3 and 4 showed normal hepatocytes. These findings suggest that annona muricata leaf extract may have both tissue protective and antioxidant properties.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Photoaffinity‐Mediated Identification of a Third Citalopram Analog Binding Site on the Serotonin Transporter
Authors: Tomlinson Michael J. , Kumar Vivek , Newman Amy H. , Lever John R. , Henry L. Keith , Vaughan Roxanne A. 
Published: June 2021

In the CNS, serotonin (5HT) controls anxiety and mood, and disorders caused by imbalanced 5HT include Major Depressive Disorder and Autism Spectrum Disorder. The serotonin transporter (SERT) is a major regulator of synaptic 5HT levels and is the target for antidepressants such as escitalopram (S‐CIT) and fluoxetine, which bind to and inhibit the transporter to elevate free transmitter. The efficacy of these drugs is moderate and many patients experience inadequate relief, indicating the need for better understanding of mechanisms or development of novel therapeutics. Antidepressant effects on SERT are complex, as many induce high‐affinity actions at the S1 orthosteric site, as well as allosteric modulation of S1 function through low‐affinity binding to a distinct S2 domain. To examine S‐CIT mechanisms we developed two novel photoaffinity ligands, [125I]VK 03‐83 and [125I]VK 03‐51, that possess identical S‐CIT pharmacophores but have cross‐linking moieties appended to different positions on the citalopram core. Our findings show that both ligands bind to SERT, inhibit 5HT transport, and allosterically regulate S1 function, but display distinct pharmacological characteristics that indicate differential binding mechanisms. [125I]VK 03‐83 labeling of SERT is fully inhibited by 5HT and typical SERT inhibitors, with an S‐CIT Ki of ~1 nM, consistent with binding to S1. In contrast, although [125I]VK 03‐51 labeling is fully displaced by the homologous ligand, it is inhibited only ~30% by S‐CIT or other standard SERT inhibitors, and shows no displacement by 10 μM 5HT, indicative of binding to multiple sites. The S‐CIT‐displaceable portion of [125I]VK 03‐51 labeling occurs with a Ki of ~1 nM, consistent with binding at S1, whereas the fraction of labeling that is not sensitive to ≥10 μM concentrations of S‐CIT, other inhibitors, or 5HT likely does not occur at either S1 or S2, which should both be fully occupied by these conditions. These findings suggest that [125I]VK 03‐51 interacts with SERT at a third site that has not been previously detected by reversible binding or co‐crystallization approaches, but is revealed by covalent cross‐linking. This site may function in transport inhibition or allosteric regulation, and suggests a domain for exploration of transporter mechanisms or medication development.Support or Funding InformationNIH DA027845 and UNDSMHS Seed Grant (LKH and RAV), NIDA Intramural Research Program (AHN)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ACETAZOLAMIDE INCREASES LOCOMOTION, EXPLORATORY BEHAVIOR, AND WEIGHT LOSS FOLLOWING SOCIAL STRESS: A TREATMENT FOR EMOTIONAL EATING?
Authors: Student Jeff , Woodman Ryan , Lockette Warren 
Published: June 2021

BackgroundAmphetamines and sympathomimetics (e.g. phenylpropanolamine and ephedrine) are effective, centrally acting drugs that induce weight loss through their potent anorexic and locomotor properties. We first reported that amphetamines and these sympathomimetic agents antagonize catecholamine‐dependent, alpha‐2 adrenergic receptor‐dependent signal transduction mediated by chloride/bicarbonate transport.HypothesisOther drugs that target and inhibit cellular bicarbonate/chloride antiport would similarly demonstrate anorectic properties, induce locomotion, and diminish weight gain ‐‐especially in disorders such as depression or social isolation where there is an increase in emotional eating.MethodsMale and female inbred mice were housed in groups or they underwent the stress of living in social isolation or in solitary metabolic chambers for five days. Mice consumed either normal rodent chow or a rodent food supplemented with high fat, high fructose corn syrup; water was provided ad libitum. To inhibit chloride/bicarbonate transport, acetazolamide (ACT, 3 mM) or vehicle was added to the drinking water. There were no noticeable adverse effects in mice receiving ACT. Following a period of habituation, rodents underwent evaluations of exploratory locomotion and learning with the object recognition test. Food intake and weights were also tracked; for all groups, n ≥ 9.ResultsCompared to the control, unstressed, group housed rodents, living in social isolation reduced spontaneous exploration time in male mice (values expressed as means in sec ± SEM) from 23 ± 4 to 14 ± 3 (p<0.001), and in female mice, from 47 ± 6 to 23 ± 3 (p<0.001). Drinking water supplemented with ACT had no effect on exploration time in group housed mice, but ACT completely restored the diminished exploration time found in mice stressed by isolation to control, grouped mice values. The ratio of time spent exploring new objects compared to familiar items (DR ratio) was also reduced following social isolation from 2.6 ± 0.5 to 1.2 ± 0.2 (p<0.001). ACT completely normalized the DR ratio of the stressed mice. The body weight of female mice fed high fat, high fructose corn syrup diets for 12 weeks and given ACT was significantly reduced when compared to mice drinking only water (weight in g ± SEM), 23.7 + 0.8 v. 21.0 + 0.5, p=0.02. This difference in weight was not due to changes in body water volume, but instead, due to the significant decrease in food consumption and greater locomotor activity recorded in mice fed the ACT.ConclusionDrugs that inhibit chloride/bicarbonate transport could offer a safe, new approach to achieving behavioral weight loss in obese individuals with emotional eating.Support or Funding InformationUniversity of MissouriThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activated Protein C‐mediated Crosstalk Between PAR1 and S1PR1 in Endothelial Barrier Stabilization
Authors: Molinar Olivia , Chen Buxin , Grimsey Neil J. , Lin Huilan , Coronel Luisa Julieta , Trejo JoAnn 
Published: June 2021

Vascular leakage is a hallmark of sepsis, a life‐threatening condition induced by systemic microbial infection. Mortality rates for severe sepsis are high, due to the lack of available effective treatments. The aim of this study is to define the key signaling pathways that promote endothelial barrier stabilization to identify new cellular targets that can be manipulated to reduce vascular leakage. Activated protein C (APC) is an anti‐coagulant protease that was previously shown to promote endothelial barrier protection through protease‐activated receptor‐1 (PAR1), a GPCR. Activation of PAR1 by APC stimulates Rac‐1 activity, which enhances endothelial barrier stability through modulation of the actin cytoskeleton and adherens junction integrity. In addition, PAR1 appears to require the sphingosine 1‐phosphate receptor‐1 (S1PR1) to promote endothelial barrier stabilization, but how signaling by the two receptors integrate to promote endothelial barrier stability is unknown. We previously demonstrated that APC/PAR1‐mediated endothelial barrier protection requires compartmentalization in caveolae and is dependent on β‐arrestin‐2 and dishevelled‐2 (Dvl‐2) scaffolds. However, it is unknown whether β‐arrestin‐2 and Dvl‐2 are required for PAR1 and S1PR1 signaling crosstalk via APC. Here, we report that PAR1 and S1PR1 co‐localize in endothelial cells and co‐segregate in caveolin‐1 enriched fractions, consistent with PAR1 and S1PR1 signaling from caveolae. We found that APC treatment of endothelial cells results in SK1 translocation to membrane fractions and activation. In addition, we show that loss of β‐arrestin‐2 expression causes a significant loss in APC‐induced SK1 activation. Thus, we hypothesize that endothelial barrier stabilization induced by APC/PAR1 and S1PR1 is integrated by β‐arrestin‐2 and SK1.Support or Funding InformationOlivia Molinar is an Institutional Research and Academic Career Developmental Award (IRACDA) Postdoctoral Fellow and is supported by NIGMS/NIH award K12GM068524. This work is funded by two “GPCR CYTOPROTECTIVE SIGNALING MECHANISMS” R01 grants: 3R01GM116597‐01S1 and 5R01GM116597‐02.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the Impact of Altered Subunit Expression on Sweet Taste Receptor Surface Trafficking and Signaling
Authors: Tan Susan , Healey Robert , Thordarson Pall , Finch Angela 
Published: June 2021

The sweet taste receptor is a heterodimeric G protein‐coupled receptor (GPCR), consisting of the subunits Tas1R2 (T1R2) and Tas1R3 (T1R3). Expression of this receptor has been demonstrated in various tissues throughout the body, including the pancreas, small intestine and adipose tissue. This receptor is thought to play a role in maintaining the body's energy homeostasis, and may potentially be a target in the treatment of metabolic disorders such as diabetes and obesity. Some studies show that the two sweet taste receptor subunits are expressed in unequal quantities in pancreatic β cells and adipocytes, however the impact of this unequal expression on receptor function is not fully understood. This study aims to determine the impact of varying the degree of expression of the two subunits on receptor surface trafficking and downstream signaling pathways. To address this aim, sweet taste receptor‐expressing stable cell lines were generated in AD293 cells by sequential selection of vectors, and in HEK293 TRex cells using bicistronic vectors and the FlpIn system. Receptor expression was characterized through real‐time PCR and biotinylation pull‐down experiments, while receptor signaling was determined in live cells using a genetically encoded Fluorescence Resonance Energy Transfer (FRET) sensor and a Bioluminescence Resonance Energy Transfer (BRET) sensor. Our results show that expression of both receptor subunits is predominantly intracellular, and that co‐expression of both subunits does not improve surface expression. Co‐expression of both subunits was, however, found to be necessary for signaling, as cell lines expressing only one subunit did not show functional responses to aspartame. The magnitude of these functional responses was found to be greatest when subunit expression was closest to 1:1, suggesting that the sweet taste receptor is most efficacious as a 1:1 heterodimer. Cell lines expressing both subunits were found to signal through Gi, as well as undergo extracellular signal‐regulated kinase (ERK) phosphorylation, demonstrating the ability of this receptor to couple to multiple signaling pathways. While some studies using mouse sweet taste receptors have reported Gs signaling at high concentrations of sweeteners, this could not be replicated in this study using human receptors, suggesting that signaling pathways may be species‐specific. Overall, this study shows that co‐expression of both sweet taste receptor subunits is necessary for signaling but not for surface trafficking and that this receptor is most efficacious when both subunits are expressed at equal levels.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the Effects of E‐Cigarette Vapor on Oral Epithelial Cells in a Cultured Cell Model
Authors: Duggar Marygrace , Swanson Hollie , Hill‐Odom Miriam 
Published: June 2021

E‐cigarette usage is growing, especially among adolescents and young adults. This trend is especially concerning due to the lack of regulations governing the e‐cigarette industry and lack of scientific data on the effects of e‐cigarettes on human health. Using a cultured oral keratinocyte model, we hypothesized that exposure to e‐cigarette vapor would adversely affect the growth of cells that reside in the oral cavity. To test this hypothesis, we cultured the cells with condensate prepared from the vapors of grape flavored e‐cigarette fluids, both with and without nicotine. After three days of treatment, cell numbers were determined using a colorimetric assay. Results indicate that the e‐cigarette condensate adversely affects cell growth, regardless of the absence or presence of nicotine. To understand the underlying mechanisms, changes in gene expression in the cells treated with condensate were evaluated using real‐time qPCR after 24, 48, and 72 hours of treatment. A time‐dependent increase in the levels of CYP1A1 and IL8, gene targets of the aryl hydrocarbon receptor, were observed. Results obtained from similar experiments using the isolated flavoring agent, methyl anthranilate, indicate that formation of a heat‐induced byproduct of this agent may be involved in the observed changes in cell growth. Future experiments will focus on determining the mechanisms by e‐cigarette condensates impact cell growth and identifying the specific chemical byproduct mediating these effects.Support or Funding InformationFunded in part by a grant from the American Society of Pharmacology and Experimental Therapeutics for Summer Undergraduate Research.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
HRH1: A Novel GPCR Drug Target in Pancreatic Cancer
Authors: Rodriguez Alyssa , Sriram Krishna , Moyung Kevin , Insel Paul 
Published: June 2021

Patients with pancreatic cancer, in particular pancreatic ductal adenocarcinoma (PDAC), have a ~6% chance of surviving 5 years after diagnosis. Approved drugs only add weeks or months to patient lifespan. PDAC is difficult to treat, in part because the tumor often has poor vascularization and dense extracellular matrix, which yield a hypoxic tumor microenvironment. Finding new drugs for PDAC is thus an unmet medical need. The G protein‐coupled receptor (GPCR) superfamily is the most frequently targeted class of proteins for FDA‐approved drugs. We hypothesized that one or more GPCR may be a novel therapeutic target for PDAC.Bioinformatic analysis of PDAC tumors in TCGA [The Cancer Genome Atlas] revealed prominent (~32‐fold) overexpression of H1 histamine receptor (HRH1) compared to normal pancreatic tissue [GTex database]. KRAS and p53 mutations occur, respectively, in ~70% and ~60% of PDAC tumors whereas increased HRH1 expression of HRH1 in 100% of tumor samples in TCGA. HRH1 is an attractive target for repurposing since many FDA approved antagonists are directed at this receptor. We thus sought to validate HRH1 expression in PDAC cells.Using qPCR, we found high HRH1 expression and low expression of HRH2‐4 in BxPC, AsPC‐1, and CAPAN‐2 (PDAC) cell lines and normal pancreatic ductal epithelial cells. Addition of histamine increased Ca++ in ASPC‐1 and CAPAN‐2 cells; this increase in Ca++ was blocked by multiple FDA‐approved HRH1 antihistamines. Histamine increased PDAC cell proliferation in cells grown in hypoxia but not normoxia. High expression of HRH1 in PDAC cells may thus confer a growth advantage for PDAC cells in hypoxic conditions. These preliminary data suggest that FDA‐approved HRH1 antihistamines are candidates for repurposing as novel therapeutics for the treatment of pancreatic cancer.Support or Funding InformationSupported by HHMI and an ASPET David Lehr AwardThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
GPR68, a proton sensing GPCR, mediates interaction of pancreatic cancer associated fibroblasts and cancer cells
Authors: Wiley Shu , Sriram Krishna , Liang Wenjing , Chang Sarah , McCann Thalia , Nishihara Hiroshi , French Randall , Lowy Andrew , Insel Paul 
Published: June 2021

Pancreatic ductal adenocarcinoma (PDAC) has a unique tumor microenvironment that is characterized by a dense fibrotic stroma (desmoplasia) that is generated by pancreatic cancer associated fibroblasts (PCAFs) derived from pancreatic stellate cells (PSCs) and pancreatic fibroblasts (PFs). Using an unbiased GPCRomic array approach we identified GPR68, a proton sensing GPCR, has much higher expression in PCAFs compared to both PFs and PSCs. GPR68 also had increased expression in PDAC tumors compared to normal pancreas and its expression was increased in PSCs by co‐culture with PDAC cells or incubation with TNFα. GPR68 activation in PCAFs enhanced expression of interleukin‐6 (IL‐6) via a cAMP/PKA/CREB signaling pathway and GPR68 knockdown with siRNA diminished IL‐6 production by PCAFs and an enhancement in proliferation of PDAC cells by PCAF conditioned media. Use of the allosteric modulator compound ogerin enhanced pH‐dependent increase in cAMP. Furthermore, a pilot screening of 96 GPCR specific compounds tested with GPR68 overexpressing HEK cells identified five compounds that decrease intracellular cAMP at pH6.4, indicating that they may be inhibitors of GPR68. We conclude that PDAC cells induce expression of GPR68 in PCAF, resulting in increased IL‐6 production by PCAFs and enhanced PDAC cell proliferation. This PCAF‐expressed GPR68 thus contributes to PDAC cell‐PCAF interaction and may be a novel therapeutic target for pancreatic cancer, a type of cancer for which new therapies are an important unmet medical need.Support or Funding InformationThis work was supported by National Institutes of Health (NIH), National Cancer Institute (NCI) Therapeutic Training Grant 5T32CA121938, NIH/NCI Research Grants R21 CA189477, R01 CA155620, and Padres Pedal the Cause PTC2017, and by an American Society for Pharmacology and Experimental Therapeutics (ASPET)‐David Lehr Research Award.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of G protein‐coupled receptors in pancreatic cancer cells
Authors: Liu Dongling , McCann Thalia , French Randall , Wiley Shu , Lowy Andy , Insel Paul 
Published: June 2021

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal forms of cancer and has an overall 5‐year survival rate ~7%. PDAC has poor response to chemotherapy and radiotherapy; thus new treatments for PDAC are an important, unmet medical need. G protein‐coupled receptor (GPCR) regulate many biological functions, including in cancer cells, but GPCR expression and the role of GPCRs in PDAC cells is unclear. We generated two primary PDAC cell lines (79E and 34E) and an immortalized normal pancreatic ductal epithelial cell line (HPDE6). Hypothesizing that GPCRs may regulate PDAC cells, we used Taqman GPCR array to assess GPCR expression in four PDAC cell lines, 79E, 34E, Miapaca, and AsPC. We identified 68 commonly expressed GPCRs in all four PDAC cell lines, among which 22 GPCRs were significantly up‐regulated (at least 2‐fold) in PDAC cells compared to normal control cell HPDE6; 13 GPCRs were significantly down‐regulated and 3 GPCRs were uniquely expressed in PDACs. We further tested the 13 most up‐regulated GPCRs using real time qPCR and confirmed that LPHN1, CHRM4, FZD4, HRH1 and CD97 have much higher expression in PDACs compared to HPED6, suggesting that these GPCRs might contribute to the malignant phenotype and perhaps could be novel treatment targets for pancreatic cancer, in particular for PDAC.Support or Funding InformationThis work is supported by NIH/NCI Research Grants R21 CA189477, R01 CA155620, and Padres Pedal the Cause PTC2017.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TARGETING THE METABOLIC SIGNALING NETWORK FOR IMMUNE CELL LONGEVITY AND FUNCTION IN ORAL CANCER THERAPY
Authors: Allevato Michael 
Published: June 2021

Oral squamous cell carcinoma (OSCC), which includes cancer of the oral cavity and oropharynx, is a significant health issue. Despite aggressive multimodality therapies, OSCC has a poor five year survival rate (60%), which results in ~300,000 deaths each year worldwide1. OSCC is associated with human papillomavirus (HPV) infection, and abusive intake of tobacco and alcohol 2–3. OSCC evades the immune recognition and anti‐tumor immune response, by exploiting the immune system's regulatory immune suppressive mechanisms 2–3. A key mechanism of tumor immune suppression has been depicted as T‐cell exhaustion 4–7. In this case, T‐cell's effector function is restrained due to the activation of checkpoints, such as PD‐1 by its ligand PD‐L1, expressed on cancers cells, T‐cells, natural killer cells and macrophages 4–7. Immune checkpoint inhibitors have demonstrated revolutionary restoration of T‐cell mediated anti‐tumor immunity in OSCC 4–7. Despite this success, the overall response rate of these immune therapies in OSCC is only ~20 percent 4–7. New therapeutic options to prevent and treat OSCC are urgently needed. The unrestrained growth of cancer cells and the effector function of immune cells are often supported by the same metabolic pathway, namely aerobic glycolysis8. We believe that by taking advantage of the unique metabolic requirement of tumor cells and the cancer‐fighting immune cells, we can discover new insights into how such a relationship regulates anti‐tumor immunity. Our aim is to elucidate the role of immune and cancer cell metabolic properties in regulating the efficacy of the anti‐tumor response. Targeting both the metabolic signaling pathways of the OSCC tumor cells and tumor infiltrating immune cells has the potential to achieve cancer remission by both halting OSCC progression and enhancing the longevity of the anti‐tumor immune response.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.