bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterizing Insulin Sensitivity in Adult LEW.1WR1 Rats
Authors: Mercado Luis Daniel , Collins Genoah , Clopp Amelia , Love‐Rutledge Sharifa 
Published: June 2021

Diabetes prevention is a large topic of research that is focused on nutritional and exercise‐based interventions with some genetic predisposition based interventions. FAT10 is a ubiquitin‐like protein and Type 1 Diabetes (T1D) susceptibility gene that may play a role in age‐related inflammation, adiposity, cancer, and kidney disease. The young T1D susceptible, LEW.1WR1 rat overexpresses FAT10 and has higher fasting concentrations of blood insulin. It is unclear how the overexpression of FAT10 effects directly affects insulin sensitivity, secretion, or production. It is also unclear if the initial insulin sensitivity of this animal model plays a role in disease susceptibility. Therefore we hypothesized that older LEW.1WR1 rats will have markers of increased insulin resistance like increased insulin and beta cell mass due to the poor regulation of FAT10 expression in this model. The objective of this study was to characterize insulin sensitivity, insulin levels, and characterize the relative beta cell size of adult LEW.1WR1 rats with or without dietary stress to understand if FAT10 expression plays a role in the regulation of inflammation and aging‐induced insulin resistance. To test this hypothesis we used LEW.1WR1 and LEW.SsNHsd rat and monitored weight gain and glucose tolerance over the course of 12 weeks on a high fat or control diet; we also isolated pancreata and used immunohistochemistry staining for insulin to analyze islet size and insulin staining. The LEW.ssNHsd rat served as a control rat because it does not have the alteration in the FAT10 promoter region. Glucose tolerance tests were performed prior to starting the diet as a baseline, and after 10 weeks on the diet to determine the effects of diet and aging on the rat groups in regards to Insulin Sensitivity. We observed that in the course of 10 weeks the control LEW.1WR1 rats became significantly more glucose intolerant and had increased insulin levels. The control LEW.1WR1 rats also gained weight at a rate similar to the high‐fat diet, a diet that is traditionally used to induce insulin resistance and glucose intolerance. This data suggests that FAT10 may be playing a role in age‐related adiposity, glucose intolerance, insulin sensitivity, and beta‐cell changes in the LEW.1WR1 rat. This study begins to lay the groundwork for understanding how alterations in FAT10 and metabolism increase T1D susceptibility.Support or Funding InformationThe University of Alabama in HuntsvilleLouis Stokes Alliance for Minority Participation (NSF)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Noncanonical Role of Delta Protein Kinase C (δPKC) Phosphorylation of Troponin I in Cardiac Reperfusion Injury
Authors: Lin Amanda J. , Ostberg Nicolai P. , Kornfeld Opher , Qvit Nir , Mochly‐Rosen Daria 
Published: June 2021

Injury of the myocardium due to reperfusion following an ischemic period (ischemia/reperfusion injury; IR) is the consequence of an imbalance between the production of oxidants and the availability of endogenous antioxidants. Depriving the heart of oxygen and nutrients (ischemia) leads to impaired mitochondrial function, elevated mitochondrial fission, cardiac damage, and death. Paradoxically, restoring blood flow (reperfusion) to the heart following an ischemic event poses another type of insult in the form of increased mitochondrial dysfunction, apoptosis, and necrosis, resulting in cardiac failure. Our laboratory has found that δPKC mediates reperfusion injury; treatment with a selective inhibitor of δPKC, δV1‐1, at reperfusion reduces infarct size by 60% in a variety of animal models. One of several substrates of δPKC, cardiac troponin I (cTnI), undergoes partial proteolysis upon phosphorylation by δPKC during IR. Interestingly, using western blot and microscopy studies, we found that the N‐terminal fragment of cTnI co‐localizes with mitochondria following IR and that inhibition of δPKC by δV1‐1 inhibits this cleavage. To determine the role of phosphorylated cTnI, we rationally designed a highly selective inhibitor of cTnI phosphorylation by δPKC, ψTnI. ψTnI inhibits only cTnI phosphorylation and not the phosphorylation of pyruvate dehydrogenase kinase (PDK), myristoylated alanine rich C kinase substrate (MARCKS), signal transducers and activators of transcription (STAT), and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) – substrates of δPKC that are elevated during reperfusion injury to the myocardium. ψTnI also diminishes cTnI association with mitochondria following reperfusion. Importantly, ψTnI treatment alone was sufficient to obviate acute IR injury in a whole heart ischemic model, reducing infarct size by over 70% and improving mitochondrial respiration when given at reperfusion. Additionally, ψTnI treatment at reperfusion of H9c2 cardiomyocytes restored mitochondrial oxygen consumption, preserved mitochondrial morphology, and rescued mitochondrial membrane potential in a dose‐dependent manner. We will determine the effects of cTnI phosphorylation and fragment association with mitochondria using loss‐ and gain‐of‐function mutations, live‐imaging, and functional assays. Together, our study identified a non‐contractile role for cTnI; in addition to regulating cardiac contractility, we find that cTnI regulates mitochondrial function following reperfusion injury to the heart.Support or Funding InformationNIH grant R01‐HL052141 to DM‐R and AHA Postdoctoral Fellowship 19POST34380299 to AJLThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protective Effect of Intranasal Peste des Petits Ruminants Virus and Bacterine Vaccinations: Clinical, Haematological, Serological and Serum Oxidative Stress Changes in Challenged Goats
Authors: Emipke Benjamin Obukowho , Jarikre Theophillus Aghogho , Jeremiah Olalekan Adetokunbo , Akpavie Stephen Adetokunbo 
Published: June 2021

The current vaccination for Peste des Petits ruminants PPR is stalled by myriad challenges and continuous endemicity of pneumonia due to fulminant bacteria complication in goats. The present study evaluated the protective effect of intranasal PPR virus linage 1 and bacterine vaccinations. Twelve West African Dwarf goats aged 6 months were randomly grouped and vaccinated within two weeks using combination of PPRV linage 1 vaccine (Nig/75), and bacterine from Mannheimia haemolytica (Mh) or Pasteurella. multocida (Pm) intranasally as A (vaccinated with PPRV & Mh), B (vaccinated with PPRV & Pm), and C (unvaccinated). All the goats were observed for three weeks post vaccination before comingling with a known infected WAD goat showing clinical signs of PPR and further observed clinically for five weeks (35days) post infection. Blood samples were taken for haematology and while sera were assayed for antioxidants (Glutathione Peroxidase GPx, Glutathione Transferase GST, Superoxide Dismutase SOD) activities and prooxidants (Malondialdehyde content, Reduced Glutathione GSH, Hydrogen peroxide H2O2 generation, Myeloperoxidase) using spectrophotometric methods. Data was subjected to parametric statistics at α=0.05 using Graph pad prism version 21. Clinically, there was pyrexia, mucopurulent oculo‐nasal discharge, diarrhoea, anaemia, leucopaenia and increased prooxidants in the unvaccinated goats (C), while moderate neutrophilia and leucocytosis were observed in PPRV and bacterine vaccinated goats. Two unvaccinated goats died at 13 and 28 days post infection. The goats vaccinated with PPRV and Mh showed better response. In conclusion, mucosal vaccination of goats with PPRV vaccine and bacterine will protect against exposure and culminate in the development of protective mucosal, humoral and cell‐mediated immune responses. This vaccination strategy will provide protection needed in the prevention and control of endemic caprine pneumonia in Nigeria.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
“Conditional Deletion of Hepatocellular Integrin Linked Kinase (hILK) Promotes an Increase in Hepatic Phosphoinositide 3‐kinase delta (PIK3Cδ)”
Authors: Martucci Nicole Jenna , Cotner Mary‐Clare , Bhushan Bharat , Mars Wendy M , Michalopoulos George K 
Published: June 2021

IntroductionBiliary diseases accounted for over 100,000 deaths in the United States in 2016. When biliary cells (cholangiocytes) experience chronic injury during these diseases, hepatocytes can transdifferentiate into cholangiocytes to supplant the lost biliary cells. While the phenomena of transdifferentiation in the liver has been observed by many, the proteins and signaling pathways involved are not yet widely understood. Data from our lab has shown that with hepatocyte specific knockout of Integrin Linked Kinase (hILK KO), there is hepatocyte proliferation, increased matrix deposition, and an unorganized biliary cell/ductal proliferation; this led us to hypothesize that removal of hepatocellular ILK induces transdifferentiation of hepatocytes to cholangiocytes.MethodsWe originally generated hILK KO mice by repeatedly crossing mice expressing cre recombinase under an afp‐albumin promoter (Cre+/−) with mice that have floxed ILK genes (ILKfl/fl) to obtain mice that were ILKfl/fl:Cre+/− (hILK KO) and ILKfl/fl:Cre−/− (WT). To identify RNAs potentially involved in transdifferentiation, we performed data array analyses on livers from 7‐ and 14‐week old hILK KO and WT mice. RNAs of interest were then further examined by Western blot analyses and immunohistochemical staining.ResultsAs anticipated from our previous published results, at 14 weeks, our data uncovered specific changes in RNAs related to matrix composition. A significant decrease was observed in Syndecans 1, 2 and 4 in the hILK KO mice. Collagen composition also changed as Col3a1, Col4a1, and Col1a4 and Col1a2 all increased in the hILK KO animals. Unexpectedly, our 7‐week RNA data analyses suggested a central role for Phosphoinositide 3‐kinase delta (PIK3Cδ) as it was commonly found in several of the Ingenuity Pathway Analysis pathways that were identified. PIK3Cδ is considered a cytoplasmic, leukocyte‐specific protein, however, in the hILK KO, the protein was also found in the nuclei of cholangiocytes and hepatocytes. pAKTS473, which is known to be downstream in PI3 kinase signaling pathways, and typically known as a plasma membrane protein, was also found in these nuclei, suggesting a direct relationship. We hypothesize nuclear PIK3Cδ is involved in the transdifferentiation of hepatocytes to biliary cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
NF1 dissociates cell type specific contributions toward reward valuation and motor control
Authors: Sutton Laurie , Dao Maria , Brian Muntean , Martemyanov Kirill 
Published: June 2021

Neurofibromin 1 (NF1) is a large multidomain signaling molecule that is a major Ras regulator but also acts as a positive regulator of cAMP levels by mediating G protein‐coupled receptor (GPCR)‐dependent activation of adenylate cyclase. Mutations in the NF1 gene cause Neurofibromatosis type 1, a genetic disorder characterized by multiple benign and malignant tumors with prominent neuropsychiatric symptoms that include learning and attention deficits, as well as motor impairments. Recently, we found that NF1 is a direct effector of GPCR signaling via Gβγ subunits in the striatum. However, it remains unclear the signaling contribution of NF1 in striatal‐mediator behaviors. Here we demonstrate the impact of intracellular signaling pathways commonly targeted by neurotransmitter inputs that impact medium spiny neurons (MSN) activity can be dissociated in a cell‐specific manner with distinct contribution to motor learning and reward‐related behaviors. We genetically ablated NF1 with cellular resolution demonstrating the diverging behavioral and signaling specific manner in which NF1 contributes to the acquisition and consolidating of learning a motor skill, as well as its regulation of morphine reward.Support or Funding InformationThis work was supported by the NIH and by the Canadian Institutes of Health Research (CIHR) Fellowship.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Establishing a time course for spontaneous opioid withdrawal in male and female outbred mice using an abbreviated dependence paradigm
Authors: Quock Raymond Mark , Brewer Abigail Lucille 
Published: June 2021

Opioid withdrawal is often a limiting factor in recovery from opioid dependence due to negative reinforcement caused by continued use of opioids in order to avoid the distressing withdrawal symptoms. In animal models of dependence, opioid withdrawal is frequently precipitated using opioid antagonists and the severity of withdrawal is related to the level of dependence. However, this model lacks face validity when investigating new treatment possibilities for opioid withdrawal because most human addicts are not exposed to an antagonist in order to precipitate withdrawal. Furthermore, results from animal experiments suggest that spontaneous and precipitated withdrawal involve different mechanisms and may favor different symptoms of withdrawal. These experiments were conducted to investigate the timeline of spontaneous withdrawal in an outbred mouse model using an abbreviated dependence paradigm. Male and female NIH Swiss mice were injected with escalating doses of morphine twice daily (50–125 mg/kg morphine) over a period of four days. On the fifth day, mice were given a final dose of morphine and withdrawal symptoms were assessed at 5, 10, 24, 34, 48, 58, 72, 82, 96, and 106 hrs after the final injection. Withdrawal symptoms were assessed by global scores for an overall picture and by counting the number of tremors, wet dog shakes, and jumps. Results showed a robust withdrawal time course that peaked at 24 hrs in both sexes as assessed by global score. When broken down, wet dog shakes and tremors proved to be the most reliable indicator of severity of withdrawal and females showed more tremors than males for the 1st 24 hours of the time course. These results establish a spontaneous withdrawal time course in an outbred strain of mouse that shows that dependence can be reliably established over a four‐day period using fairly high escalating doses of morphine. Furthermore, these results suggest that individual symptoms of withdrawal should be assessed over time as well as global score.Support or Funding InformationSupported in part by the State of Washington Initiative Measure No.171.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Novel sumanirole bivalent analogues as potent dopamine D2 receptor Go‐protein biased agonists
Authors: Yano Hideaki , Bonifazi Alessandro , Hoffman Alexander F , Lupica Carl R , Shi Lei R , Newman Amy H 
Published: June 2021

The dopamine D2 receptor (D2R) is a major target for therapeutic development of treatments for neuropsychiatric disorders. In particular, D2R agonists have been used for neurological disorders such as restless leg syndrome and Parkinson's Disease. However, current D2R agonist pharmacotherapies have drawbacks including the development of tolerance and other adverse side‐effects. To potentially improve D2R agonist‐based medications, we have designed bivalent ligands that have two pharmacophores tethered by a link. This strategy aims to improve selectivity, affinity, and cell signaling activation bias. Indeed, it has been demonstrated that the secondary pharmacophore plays a key role in inducing activation bias towards either G protein, or β‐arrestin pathways.Here we characterized a series of bivalent orthosteric‐arthosteric heteroaryl pharmacophores with assorted linkers and compared them with the potent orthosteric D2R‐selective agonist, sumanirole. Several bivalent analogues demonstrated modest increases in D2R affinity compared to sumanirole. D2R functional assays using bioluminescence resonance energy transfer (BRET) showed improved efficacy and potency for G protein coupling, but substantially diminished efficacy and potency for β‐arrestin recruitment. Analysis using an operational model of bias revealed significant G‐protein activation bias (>1000‐fold). Further, several compounds exhibited a G protein subtype (GoA over Gi1) activation bias. Finally, D2R‐mediated, G protein‐coupled inwardly‐rectifying potassium channel (GIRK) activity was measured in midbrain slices using electrophysiology. Recordings from D2R‐expressing dopaminergic cells in the ventral tegmental area confirmed prolonged GIRK activation by the biased bivalent compounds, compared to dopamine or sumanirole.Our novel sumanirole bivalent analogues demonstrate a proof of concept for G protein activation bias and thus may lead to D2R agonist therapeutics with reduced tolerance and improved efficacy. With these novel tools, the preferential targeting and activation of Go‐proteins, the most predominant Gi‐like protein family in the brain, are now conceivable in animal models. This advance may aid in treatments of basal ganglia disorders with reductions in untoward side‐effects in humans.Support or Funding InformationWork was supported by the Intramural funds of the National Institute on Drug AbuseThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gβ5‐R7 regulator of G protein signaling complex is a positive regulator of insulin secretion
Authors: Slepak Vladlen , Wang Qiang , Henry Taylor , Pronin Alexey 
Published: June 2021

We present new data describing the function of the atypical G protein beta subunit Gβ5 (gene: Gnb5). Gβ5 is unique in that instead of Gγ dimerizes with regulator of G protein signaling proteins of R7 family (RGS6, 7, 9 and 11). These obligatory protein complexes are known to be highly expressed in neurons, but have also been found in other tissues, for example in endocrine glands. Here, we show that Gβ5‐R7 plays a novel and surprisingly important role in stimulated insulin exocytosis. We demonstrated earlier that CRISPR/Cas9‐mediated deletion of the Gnb5 gene ininsulin‐secreting cell line MIN6 results in a dramatic reduction insulin secretion induced by muscarinic M3 receptor (M3R). Our data on the knockout of Gnb5 in mice also results in about 2‐fold lower level of serum insulin, and in a dramatic reduction in M3R‐stimulated secretion occurs in Gnb5−/− primary islets. These results are unexpected because in other systems, the knockout of this RGS protein complex facilitates signal transduction, apparently to its GAP activity toward Gi family proteins. In this study, we show that deletion of Gβ5‐R7 strongly attenuates inputs from different secretagogues, not only M3R. The knockout of the Gnb5 gene affects Gq‐ and Gs‐mediated stimulation, but most surprisingly, there is a significant effect on glucose, the permissive factor for insulin release. The Gnb5 knockout does not impair second messengers cAMP, Ca2+and DAG in beta cells, but rather causes changes protein phosphorylation pattern. Our experiments also implicate regulation of some protein kinases including ERKs and ROCK and participation of cell‐to‐cell interaction mechanisms, possibly adhesion GPCRs. Thus, our experiments point to the existence of a novel function of the Gβ5‐R7 complex that may be not directly associated with its interaction with heterotrimeric G proteins.Support or Funding InformationThis work was supported by U.S. National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases Grants R01DK105427 (to V.Z.S.) and R01DK111538 (to Alejandro Caicedo). The authors declare no conflicts of interest.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Parathyroid‐targeted overexpression of Regulator of G‐Protein Signaling 5 (RGS5) causes hyperparathyroidism in transgenic mice
Authors: Balenga Nariman 
Published: June 2021

The relationship between impaired calcium sensing, dysregulated parathyroid hormone (PTH) secretion, and parathyroid cell proliferation in parathyroid neoplasia is not understood. We previously reported that a GTPase activating protein, regulator of G‐protein signaling 5 (RGS5) is overexpressed in a subset of parathyroid tumors associated with primary hyperparathyroidism (PHPT), and that RGS5 can inhibit signaling from the calcium‐sensing receptor (CASR). In vivo, we found that RGS5‐null mice have abnormal PTH secretion. To gain a better understanding of the potential role of RGS5 overexpression in parathyroid neoplasia and PHPT and to investigate whether inhibition of CASR signaling can lead to parathyroid neoplasia, we created and characterized a transgenic mouse strain over‐expressing RGS5 specifically in the parathyroid gland. These mice develop hyperparathyroidism, bone changes reflective of elevated PTH, and parathyroid neoplasia. Further, expression of exogenous RGS5 in normal human parathyroid cells results in impaired signaling from CASR and negative feedback on PTH secretion. These results provide evidence supporting the setpoint hypothesis of parathyroid neoplasia and define a role for RGS5 in the pathogenesis of PHPT through inhibition of CASR signaling.Support or Funding InformationPhRMA Foundation and American Cancer Society research grants.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Remodeling the interactions between TDP43 and RNA for development of therapeutics for ALS
Authors: Felemban Razaz A , François‐Moutal Liberty , Sayegh Melissa , Perez‐Miller Samantha , Gokhale Vijay , Zarnescu Daniela C. , Khanna May 
Published: June 2021

Developing effective treatment strategies for amyotrophic lateral sclerosis (ALS), that affect upper and lower motor neuron, requires an understanding of the underlying cellular pathway that lead to motor neuron death and muscle atrophy. RNA dysregulation is a hypothesized disease mechanism in ALS. Studies have shown that pathological transactive response (TAR) DNA Binding Protein (TDP‐43) binds irreversibly to RNA in stress granules and forms membraneless prion‐like TDP‐43 aggregates in cytoplasmic motor neurons1. These membraneless compartment can form in cells through liquid‐liquid phase separation (LLPS). Recent study showed that mRNA secondary structure promotes LLPS to build membraneless compartments in cells2. Based on the TDP‐43. mRNA complex remodeling hypothesis by Coyne et al3, we hypothesized that remodeling the interactions between TDP43 and RNA using small molecules will decrease motor neuron toxicity. Using in silico docking, we screened 50,000 compounds on the RNA recognition motifs 1 and 2 of TDP43 (RRM1 and RRM2). We identified rTRD001 that was able to bind to TDP43 in RRM1 domain, disrupt TDP43 interaction with RNA, reduce cytoplasmic aggregate and cytotoxicity in motor neuron cell line NSC34, and mitigate TDP‐43 dependent phenotypes in an ALS fly model of TDP‐43 proteinopathy. Further studies are necessary in order to assess the mechanism and mode of action for this neuroprotective effect for this compound.Support or Funding Information‐This project was funded by a grant from NIH R01 NS091299 and ABRC/ADHS16‐162407 ‐ Scholarship funded by King said bin Abdulaziz university for health science, Jeddah, Saudi ArabiaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.