bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulators Of G‐Protein Signaling 2 And 4 Differentially Regulate the Rewarding Effects of Nicotine and Cocaine
Authors: Emerson Nathaniel , Asswini Sara , Rose Madison D. , Seeley Sarah , Rorabaugh Boyd R. , DSouza Manoranjan S. 
Published: April 2020

Addiction to nicotine and cocaine is a major cause of preventable premature morbidity and mortality. The reinforcing effects of nicotine and cocaine are primarily mediated by increasing dopamine in the mesolimbic reward pathway. The effects of neurotransmitters like dopamine in this pathway are mediated by G protein‐coupled receptors (GPCRs). Interestingly, regulator of G protein‐signaling (RGS) proteins negatively regulate the functioning of GPCRs. In this study we specifically focused on the role of RGS2 and RGS4, which are extensively distributed in the mesolimbic reward pathway. This study evaluated the effects of RGS2 and RGS4 in the rewarding effects of nicotine (0.5 mg/kg; s.c.; base) and cocaine (15 mg/kg; i.p.) using conditioned place preference procedure (CPP) in mice lacking either RGS2 (i.e. RGS2 KO) or RGS4 (RGS4 KO) and their wildtype counterparts. Because a role of sex has been reported in the effects mediated by different RGS proteins, both male and female mice lacking either RGS2 or RGS4 were studied while assessing the rewarding effects of cocaine and nicotine. Cocaine and nicotine‐induced CPP was observed in all wildtype mice irrespective of genotype and sex. Cocaine‐induced rewarding effects were attenuated in male RGS4 KO mice, but not male RGS2 KO mice compared to respective saline controls. In contrast, nicotine‐induced rewarding effects were attenuated in male RGS2 KO mice, but not in male RGS4 KO mice compared to respective saline controls. Interestingly, knockout of either RGS2 or RGS4 in female mice had no effect on nicotine or cocaine‐induced rewarding effects, respectively. Thus, these data suggest both genotype and sex‐dependent differential effects of RGS2 and RGS4 in the rewarding effects of nicotine and cocaine. Clinically, the data support a role for targeting RGS2 and RGS4 for the treatment nicotine and cocaine addiction, respectively.Support or Funding InformationThis work was supported by a Bower, Bennett and Bennett grant awarded to Dr. Manoranjan S. D’Souza by the Raabe College of Pharmacy, Ohio Northern University (ONU), Ada, Ohio.
Bone Morphogenetic Protein (BMP) Transcripts in Kidney Tissue of Cats with Renal Dysfunction
Authors: Panickar Kiran S. , Tavener Selena , Hollar Regina L. , Jewell Dennis E. 
Published: April 2020

Bone morphogenetic proteins (BMPs) are growth factors that belong to the transforming growth factor‐β (TGF‐β) superfamily, and till date 15 BMPs have been described. BMPs, first described for their role in bone and cartilage formation, also play a role in renal fibrosis in chronic kidney disease (CKD). There is evidence to indicate that in rodent models of CKD, administration of recombinant BMP1–3 increases renal fibrosis whereas administration of a BMP1–3‐neutralizing antibody or BMP‐7 antibody reduces renal fibrosis and preserves renal function. The aim of the present study was to investigate changes in gene expression in the renal cortex obtained from cats with kidney disease or calcium oxalate stone formers (CaOx) at necropsy, to identify BMPs associated with renal dysfunction in cats and potential fibrosis. At time of death the circulating levels of creatinine as well as symmetric dimethyl arginine (SDMA), both markers of kidney decline in cats, were significantly higher in cats with renal disease (n=11) or stone‐forming cats (CaOx, n=12) when compared to controls (n=19). Using RNAseq in kidney tissue, we found a modest, but significant, increase in the expression of BMP‐1 in cats with kidney disease (2.48 fold) and stone formers (1.72 fold), compared to controls (both p<0.01). While the increase in BMP‐2 in CaOx cats was significant (1.46 fold; p<0.05 vs Con), the increase in cats with kidney disease was not (1.23 fold; NS). BMP2K, a BMP‐2 inducible kinase, was significantly increased in both kidney disease (1.43 fold) and CaOX (1.46 fold) (both p<0.05). In contrast, a significant decrease in BMP4 was observed in both groups (<2.2 fold and 1.68 fold in kidney disease and CaOx, respectively; both p<0.001 vs Con). A decrease was also seen in CRIM 1, a protein associated with podocyte filtration function and whose reduction is associated with fibrosis, in both groups. BMP‐7, whose potential therapeutic role in treating CKD and reversing fibrosis has been documented, was modestly decreased in both groups (both less than 1.5 fold) compared to controls. Given that there was an increase in all three forms of TGFβ (TGFβ1, TGFβ2, and TGFβ3), a potent initiator of renal fibrosis, in both groups, and a decline in BMP‐7, an endogenous inhibitor of TGFβ signaling in fibrosis, compared to controls, our results profile the BMPs potentially associated with renal fibrosis in cats that may contribute to kidney dysfunction. In summary, a nutritional therapy to slow the progression of kidney dysfunction may benefit from the inclusion of dietary ingredients that attenuate renal fibrosis in cats.Support or Funding InformationThis study was funded by Hill’s Pet Nutrition, Inc
Local application of FTY720 hydrogel to the carotid body area reversed hypertension induced by obesity and leptin infusion
Authors: Shin Mi Kyung , Kim Lenise J. , Tang Winnie Wan-yee , Yeung Bonnie Ho-yee , Gu Chenjuan , Mitrut Roxana , Sham James S. K. , Cui Honggang , Polotsky Vsevolod Y. 
Published: April 2020

RATIONALEObesity increases the level of an adipocyte‐produced hormone leptin, which enhances the sympathetic activity leading to hypertension. However, the mechanism of the hypertensive effect of leptin in obesity is unclear. We have recently reported that acute leptin infusion induces hypertension via the Transient Receptor Potential Channel subfamily M member 7 (trpm7) in the carotid bodies (CB) (Shin et al., 2019). We hypothesize that this mechanism plays a role in hypertension when leptin levels are elevated chronically in diet induced obesity (DIO).METHODSWe have developed a novel technique allowing continuous sustained release of the Trpm7 inhibitor FTY720 into the mouse CB using a hydrogel formulation. (1) The effect of FTY720 on blood pressure during chronic leptin infusion was monitored by telemetry implanted in the left femoral arteries of male lean C57BL/6J mice (n=14). FTY720 (N=8) or control hydrogel (N=6) was applied locally to the CB region. Blood pressure was recorded at baseline and during leptin infusion (120 μg/day for 3 days once a week via a SC pump) over a 3 week interval after hydrogel application. (2) The effect of FTY720 on blood pressure in mice with endogenously high leptin levels was determined similarly in DIO mice (DIO, N=16). Blood pressure was measured at baseline and after a single application of FTY720 (N=10) or control hydrogel (N=6) to the CB.RESULTS(1) In C57BL/6J mice, administration of leptin increased mean arterial pressure by 12.9 mmHg (p<0.0001) at baseline and throughout the control hydrogel treatment over the 3 week interval. FTY720 hydrogel completely abolished leptin‐induced hypertension (p < 0.05). (2) DIO mice showed elevated blood pressure levels, compared to lean mice at baseline (ΔMAP=+16.2 mmHg). FTY720 hydrogel decreased blood pressure (ΔMAP = −7.8 mmHg, p<0.005), whereas control hydrogel did not have an effect (ΔMAP = −1.7 mmHg, p=0.529).CONCLUSIONSWe found that chronic hyperleptinemia induces hypertension, which can be treated by a single injection of an extended release preparation of FTY720 to the CB area. Our data (1) provide a new insight into the role of leptin signaling via Trpm7 in the CB in the pathogenesis of hypertension in obesity, and (2) suggest that injections of FTY720 hydrogel to the CB area may control blood pressure in obese patients with resistant hypertension.Support or Funding InformationHL 133100, HL 128970, 19CDA34700025
Examining the Role of the chromatin remodeler CHD5 in Tumor Suppression and Neural Differentiation in Zebrafish
Authors: Denning Ellen , Sorlien Erin , Ogas Joseph 
Published: April 2020

Gene expression and neurogenesis in vertebrates involves a wide variety of cellular machinery to coordinate the transcriptional changes that direct cellular differentiation. Critical aspects of this epigenetic machinery are chromatin remodeling factors. One such factor is CHD5, a vertebrate‐specific member of a family of ATP‐dependent chromatin remodeling proteins. CHD5 is expressed primarily in neural tissue where it is thought to contribute to neurogenesis and has been strongly linked to tumor suppression. Currently, little is known regarding the molecular mechanisms by which CHD5 contributes to neurogenesis or tumor suppression. We hypothesize that loss of CHD5 contributes to development of neuroblastoma because neural precursor cells lacking CHD5 fail to alter some aspect of their neural precursor transcriptome/epigenome. To study the biochemical properties and functional importance of CHD5 remodelers, we have developed a zebrafish model to study Chd5, the zebrafish homolog of human CHD5.Using the CRISPR/Cas9 system, we have engineered chd5 knockout zebrafish embryos. These zebrafish did not display any overt developmental phenotypes, however next‐generation sequencing of brain tissue from zebrafish embryos reveals that loss of chd5 is associated with broad changes in gene expression. These genes are both up and down regulated, suggesting Chd5 is necessary for both activation and repression of genes during brain development in zebrafish. Analysis of these genes and of their possible contribution to Chd5‐dependent phenotypes is underway.Support or Funding InformationPurdue Center for Cancer Research; Purdue College of Agriculture
Efforts Toward GPR88 Deorphanization: Challenges and Alternative Approaches to Troublesome Cell‐Based Second Messenger Assays
Authors: Elayan Abedalrahman M. , Ruddick Kristie R. , Cole Judith A. , Baker Daniel L. , Parrill-Baker Abby L. 
Published: April 2020

G protein‐coupled receptors (GPCR) comprise a family of over 800 integral membrane proteins involved in signal transduction and serve as the primary cellular sensors for chemical stimuli. About 35% of current drugs act on GPCR targets. In addition, of the 300 agents in current clinical trials, greater than 20% target novel GPCRs for which there are no approved drugs (orphan GPCRs). Ligand discovery and deorphanization efforts seeking to assign function to understudied GPCRs are not trivial. Notoriously complex in nature, GPCR‐dependent signaling centers around receptor‐ligand interactions that serve to stabilize conformational access to intracellular signaling partners, G proteins. In GPCR signaling, receptors and heterotrimeric G proteins (made of the Gα and Gβγ subunits) work together to transmit signals via downstream effectors and distinct pathways. The measurement of GPCR mediated second messengers such as cyclic AMP (cAMP), intracellular calcium (Ca2+) mobilization, and ERK activation (MAP kinase) are commonly used as tools to detect ligand induced GPCR activation. However, experimental pathway elucidation is complicated by the tendencies for pathway crosstalk, receptor desensitization, and receptor‐G protein interaction promiscuity. Cell‐based assays that monitor second messenger responses are often used as screening tools for potential GPCR agonists and antagonists, however, commonly used cell‐lines such as HEK‐293, express many endogenous GPCRs that can complicate data analysis. Many classes of Gα subunits exist: Gαs and Gα i/o G‐proteins modulate cyclic AMP (cAMP) through stimulation or inhibition of adenylate cyclase while Gαq subunits activate phospholipase C and a subsequent rapid release of intracellular Ca2+. Specifically, the orphan receptor, GPR88, has been linked with psychiatric diseases including schizophrenia and bipolar disorder. Lacking a validated endogenous agonist, the GPR88 signaling pathway remains poorly understood, however, in vivo knockout experiments and in vitro preclinical studies with small molecule synthetic agonists indicate that GPR88 couples to Gαi/o G‐proteins to inhibit adenylate cyclase activity and reduce intracellular cAMP in cell‐based assays. In this work, we describe our challenges in the study of the GPR88 signaling pathway using second messenger cell‐based assays and explain our decision to move to a real‐time NanoLuc‐based GPCR/G protein complementation assay.Support or Funding InformationNIH grant 1 R15 MH109034
Isoprenaline‐induced myocardial injury in fertile and ovariectomized female Sprague Dawley rats
Authors: Borodzicz-Jażdżyk Sonia , Czarzasta Katarzyna , Wojciechowska Małgorzata , Główczyńska Renata , Kołodzińska Agnieszka , Puchalska Liana , Opolski Grzegorz , Cudnoch-Jędrzejewska Agnieszka 
Published: April 2020

BackgroundOne of the most widely used animal model of myocardial injury is induced by injection of isoprenaline (ISO), a non‐selective beta adrenoceptor agonist, which in high doses causes oxidative stress and coronary demand‐supply imbalance in cardiac tissue.PurposeThe aim of the study was to assess the course and severity of acute ISO‐induced myocardial injury in fertile and ovariectomized female Sprague Dawley rats.Methods9‐weeks old Sprague Dawley rats were ovariectomized or sham operated, and after 3 weeks received an intraperitoneal injection with a single dose of 150 mg/kg b.w.t. ISO (ISOO and ISOP groups, respectively). Echocardiographic examination and blood collection were performed between 6 and 72 hours post ISO injection. Echocardiographic analysis included assessment of left ventricular (LV) dimensions, as well as systolic and diastolic function of LV.Moreover, the plasma concentrations of troponin I (TnI) and N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) were assessed.ResultsWe observed significantly higher LV end diastolic diameter (p<0.01), LV end diastolic area (p<0.01) stroke volume and cardiac output in ISOO when compared to ISOP group.Moreover, ISOO group showed significantly higher plasma concentrations of NT‐proBNP (p<0.001) and lower concentrations of TnI (p<0.001) in comparison to the ISOP group.ConclusionsThe course and severity of ISO‐induced myocardial injury differs significantly between fertile and ovariectomized female rats. Ovariectomy is associated with more pronounced left ventricular dilation and overload.Support or Funding InformationScientific work financed from budget funds for science in 2016–2019 as a research project of the Diamond Grant program (project No. DI2015 003045).
Dietary adaptation in lemurs, analyzed using new approaches to describing functional properties of tooth shape
Authors: Fulwood Ethan Lucas , Shan Shan , Winchester Julia , Kirveslahti Henry , Gao Tingran , Boyer Doug , Daubechies Ingrid 
Published: April 2020

The postcanine teeth of mammals are shaped by the requirements of food fracture. Tooth shape descriptors attempt to quantify those aspects of tooth form that reflect adaptation to the fracture of foods differing in material properties. Dental topography metrics, which capture features of whole occlusal surfaces using digital scans, have shown promise in classifying mammals, particularly primates, to dietary ecology. However, they suffer from potential issues arising from digital mesh processing, model overfitting, and the averaging of functional signal among tooth structures that are under potentially independent selection. Here, these issues are addressed using a recently developed dental topography metric (ariaDNE) which is less sensitive to details of mesh processing; Bayesian multinomial modelling with metrics designed to measure overfitting risk; and a tooth segmentation algorithm which allows the shapes of disaggregated tooth surface features to be quantified using dental topography metrics. ariaDNE quantifies tooth curvature using a method comparable to the originally described Dirichlet Normal Energy (DNE), but averages over local bandwidths of a tooth surface in a process that is robust to variation in tooth face count and smoothing algorithms. Combinations of dental topography metrics that included ariaDNE outperformed DNE in reclassifications of individual specimens and genera by their genus means using discriminant function analysis. Bayesian multinomial models suggest that informationally rich models including other metrics may overfit in out‐of‐sample reclassifications, however. Disaggregating teeth into regions of consistent shape did not improve predicted reclassification success, suggesting that averaging of morphological information across the tooth surface does not interfere with the ability of dental topography metrics to predict dietary adaptation. ariaDNE presents a powerful tool for the description of functional shape that can be applied to a wide range of taxa and potentially aspects of morphology across the body.Support or Funding InformationFunding was provided by the Duke Graduate School dissertation research domestic travel grant and summer research support and by the grants NSF BCS 1552848 to Doug Boyer, NSF BCS 130405 to Doug Boyer and Elizabeth St. Clair, and NSF BCS 1825129 to Doug Boyer and Arianna Harrington.
Evaluation of the Analgesic, Anti‐Cataleptic, Muscle Relaxant and Sedative Potential of The Ethanol Extract of Lophira alata Stem bark in Rodents
Authors: Iniaghe Loretta Oghenekome , Okpakpor Emma Elohor , Idemudia Joshua , Ogbuehi Joy Ozioma , Okonna Ifunanya Lydia , Ikhile Osatohamhen Favour 
Published: April 2020

BackgroundLophira alata, a woody plant found in West and Central Africa is used in the treatment of several systemic disorders. This study was carried out to evaluate some neurobehavioural properties of Lophira alata in murine models of algesia, sedation and catalepsy.MethodStem bark of Lophira alata were obtained, identified, authenticated, dried and milled to fine powder. Extraction was done using ethanol; phytochemical screening and determination of the mean lethal dose were carried out.Analgesia was evaluated using the hot plate and acetic‐acid induced mouse writhing tests at graded doses of 200–800 mg/kg while haloperidol and reserpine induced catalepsy were carried out to determine anti‐cataleptic potential of Lophira alata. Muscle relaxant activity and sedative property were assayed via the rotarod and diazepam induced sleep tests respectively.ResultsReduction in number of writhes in the acetic acid induced mouse writhing and increase in latency time on the hot plate were observed at dose levels of 800 mg/kg. Lophira alata at 400 and 800 mg/kg significantly reduced mean cataleptic score while no significant difference between control groups and Lophira alata treated animals were observed in the rotarod and diazepam induced sleep tests.ConclusionLophira alata possesses anti‐cataleptic property and reduces algesia possibly via peripherally and centrally mediated mechanisms.Support or Funding InformationNo honorarium received
A proteome‐wide assay for protein mistargeting in living cells
Authors: Lyu Ziqi , Sycks Melody M. , Genereux Joseph C. 
Published: April 2020

Nascent chains of secretory proteins are targeted to the endoplasmic reticulum (ER) to start their journey in the secretory pathway. Yet mistargeting may not be rare, especially during stress. Mistargeted nascent proteins then accumulate in the cytosol, which can be a potential threat to the proteostasis of the cell. A comprehensive understanding of how protein gets mistargeted, which kinds of proteins are more prone to mistargeting and which factors govern the regulation of mistargeting will be important. However, those questions are hard to answer due to the lack of methodologies: the gold standard relying on in vitro transcription/translation is quantitative but tedious, low through‐put and incompatible with living cells, while the naïve proxy of measuring ER maturation events is facile but not necessarily accurate. Here, we developed a protein partitioning assay by using a cytosol‐localized engineered ascorbate peroxidase (cytAPEX) or an ER lumen‐retained horseradish peroxidase (ERHRP) to proximity‐label cytosolic and ER luminal proteomes respectively. The compartmental selectivity of this assay was validated with exogenous and endogenous proteins. We showed protein localization can be independent of maturation events, like signal sequence cleavage. We measured how an ER stressor impacts the accumulation of ER proteins in the cytosol. We also characterized differential protein targeting under the condition of translocon inhibition. Using quantitative proteomics, we calculated the basal import efficiency of some ER‐directed proteins. Interestingly, these data have good agreement with the in vitro method. With this assay, we will be able to estimate the basal import efficiency or measure mistargeting at a proteome‐wide scale in living cells. This assay will also make it possible to investigate the mechanism behind mistargeting in response to ER stress.Support or Funding InformationUniversity of California, Riverside
Case Report: An Unusual Variant in Kidney Arterial Supply with Two Accessory Renal Arteries from Different Origins
Authors: Quiñones-Rodriguez Jailenne I. , Jiménez-Dietsch Sofia , Villamil-Rios Catalina I. 
Published: April 2020

Knowledge of the arterial supply of the human kidney is of special interest due to its variable nature as well as the clinical implications during renovascular interventions. This case report describes an unusual pattern in the arterial blood supply of the kidneys. The variation was discovered in an elderly Puerto Rican female during the course of a routine dissection on an embalmed cadaver. The left kidney received arterial supply from two accessory renal arteries. An upper accessory renal artery branched directly from the aorta and entered the kidney at the upper pole. A lower accessory renal artery branched from the main renal artery and entered the main part of the kidney at the level of the hilum. Additional branches appear to supply the left suprarenal gland both directly from the aorta and from the upper accessory renal artery. The right kidney was missing due to unknown causes occurring during the life of the individual.This variant is uncommon, and suggests an atypical embryological development. As the kidney ascends, it gathers its blood supply from nearby arteries, and when it reaches its final destination, one artery remains while the rest degenerate. When this process fails, multiple renal arteries are present in the adult, each with an important function that is not supplemental to that of the main renal artery. However, these extra renal arteries may represent the formation of additional blood vessels rather than an embryological vestige. Understanding these types of variations may help avoid clinical complications, especially during radiological examinations and surgical approaches, such as partial nephrectomies. This is especially true of the extra hilar artery occurring at the level of the hilum, which might be confused for a typical renal artery during imaging.