bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relationship between food intake and expression of messenger RNA encoding aromatic amino acid decarboxylase (AADC), tyrosine hydroxylase (TH), and catechol‐O‐methyltransferase (COMPT) in channel catfish.
Authors: Moyers Macy Grace , Kostner Danica , Wertz Blaine , White Taylor , Kobayashi Yass 
Published: April 2020

Obesity is linked to various metabolic disorders and associated complications, including major depressive disorder and anhedonia. How dopamine metabolism affects food intake and the development of obesity in fish, however, is not clear. Our objectives were to characterize the tissue expression of AADC, TH, and COMPT mRNA and if their expression is influenced by food intake in channel catfish. Expression of these mRNAs were examined in the brain, hypothalamus, liver, muscle, kidney, spleen, large intestine, and small intestine, as well as within different parts of the brain to examine their tissue expression profile. To study the relationship between hypothalamic expression of these mRNAs and food intake, samples were harvested at −1 hour prior to (−1), at feeding (0), and 1 hour after feeding (+1, n=4 per time point). Expression of all transcripts encoding these enzymes was detectable in all tissues examined, with increased expression in the liver. These mRNAs were more highly expressed in the hypothalamus compared to other parts of the brain examined. The expression of AADC mRNA and TH mRNA was not altered by timing of food exposure. Given that these mRNA transcripts had increased expression in the hypothalamus, the dopaminergic neural system may influence the regulation of food intake. Lack of changes of expression of these enzymes within the hypothalamus in response to food exposure may indicate that the changes in the dopamine system in response to food intake may be at the enzyme activity level rather than changes in transcription.Support or Funding InformationP20 GM103418
Increased extracellular superoxide and MMP9 attenuated COMP stabilization of BMPR2 potentially participate in pulmonary hypertension development
Authors: Alruwaili Norah , Kandhi Sharath , Kelly Melissa R. , Sun Dong , Wolin Michael S. 
Published: April 2020

The role of bone morphogenetic protein type 2 receptor (BMPR2) in the development of pulmonary hypertension was emphasized after identifying mutations of the gene encoding BMPR2 among patients with idiopathic pulmonary artery hypertension. Since then many studies were devoted to investigate restoring BMPR2 signaling as a therapeutic strategy. Also, roles for increased oxidant mechanisms have been identified in the lungs and right ventricle (RV) of several animal models of PH. However, it is unclear whether altered redox state plays a role in the regulation of the BMPR2 pathway in pulmonary vascular cells. Our current studies provide evidence for extracellular superoxide regulating BMPR2 expression via a Matrix Metallopeptidase 9 (MMP9)/cartilage oligomeric matrix protein (COMP) dependent pathway. Mice deficient of extracellular SOD (SOD3) show evidence of BMPR2 depletion both in the absence and presence of chronic hypoxia. Incubation of bovine pulmonary arteries under hypoxic conditions (1% O2 for 48h) increased MMP9 activity and expression which was associated with increased superoxide production and BMPR2 depletion. Moreover, adding exogenous superoxide dismutase (SOD) to the media inhibited MMP9 activation in response to acute hypoxia. In cultured human PASMC, 24 hours of hypoxia (1% O2) induced significant increase of MMP9 activity and reduced COMP expression. MMP9 contributes to COMP/BMPR2 depletion in chronic hypoxia based on MMP9 deficient mice showing a preserved expression of both COMP & BMPR2 in lung tissues. Moreover, MMP9 deficient mice exposed to chronic hypoxia or Sugen/hypoxia showed a marked reduction of the elevated RV systolic pressures and Fulton index measurements of right ventricular hypertrophy compared to wild type mice under the same experimental conditions. ECHO measurements of PAAT/ET ratio, which correlate inversely and linearly with mean pulmonary artery pressure, in hypoxia‐exposed MMP9 KO mice showed increased ratios compared to animals exposed to chronic hypoxia or Sugen/hypoxia. These data are consistent with depletion of MMP9 reducing the development of pulmonary hypertension. Thus, increased extracellular superoxide appears to promote pathological loop of MMP9 mediated depletion of COMP leading to destabilization of BMPR2 and loss of its pulmonary hypertensive protective actions.Support or Funding InformationSupported by NIH Grants HL115124 & HL129797
Identification, Characterization, and Treatment for a Taurine Transporter (SLC6A6) Variant Resulting in Taurine Deficiency and Pathologies in a Consanguineous Family.
Authors: Henry L. Keith , Ansar Muhammad , Ranza Emmanuella , Shetty Madhur , Paracha Sohail A. , Azam Maleeha , Kern Ilse , Iwaszkiewicz Justyna , Farooq Omer , Pournaras Constantin J. , Malcles Ariane , Kecik Mateusz , Muzaffar Waqar , Qurban Aziz , Ali Liaqat , Aggoun Yacine , Santoni Federico A. , Makrythanasis Periklis , Ahmed Jawad , Qamar Raheel , Rivolta Carlo , Sarwar Muhammad T. , Antonarakis Stylianos E. 
Published: April 2020

More than 200 Pakistani consanguineous families with multiple individuals having recessive visual impairments were evaluated by exome sequencing and genotyping. One of the families, F315, from the Kohat region of Pakistan contained 2 affected individuals with a homozygous deleterious variant Gly399Val (NM_003043.5:c.1196G>T) in the taurine transporter SLC6A6 (MIM:186854) which segregated with the phenotype of progressive retinal degeneration. Gly399 is highly conserved in vertebrates and molecular modeling suggests that the increased bulk of the valine substitution displaces Tyr138 which participates in substrate coordination and transport and is consistent with the hypomorphic variant exhibiting only ~15% of the transport capacity of the native protein as determined by taurine uptake analyses in HEK‐293 cells transiently or stably expressing SLC6A6 or the Gly399Val variant. The reduced function was attributed to a catalytic effect rather than plasma membrane levels and analysis of patient‐derived fibroblasts of affected individuals mirrored these findings. Affected individuals exhibited cone‐rod retinopathy and cardiomyopathy and remarkably taurine supplementation for 24 months resulted in resolution of the cardiomyopathy and clinical stabilization of the retinopathy.Support or Funding InformationProVisu foundation and ERC grant 219968 to S.E.A., and NIH grant DA027845 to L.K.H.
Antioxidant Activity and Phenolic Content of Chinese Balsam (Impatiens chinensis)
Authors: Oldenburg Kimberly , Henning Shelby , Soendergaard Mette 
Published: April 2020

Plants of the Impatiens genus includes more than 1,000 species. Several of the species are used as medicinal plants and are known to contain the anti‐inflammatory 2‐methoxy‐1,4‐naphthoquinone. Herbal remedies are used to treat insect bites, rashes, and poison ivy contact dermatitis, as well as management of anxiety and stress. Chinese Balsam (Impatiens chinensis) is found in North‐East India, where it is used medicinally for pain relief, promoting blood circulation, and treating urinary infections. However, very little research has been conducted to analyze the phytochemistry of I. chinensis. Here, is described the first report of antioxidant capacity and phenolic content of the seeds, leaves, stems, roots and flowers of I. chinensis.Impatiens chinensis plants were separated into seeds, leaves, stems, roots and flowers, freeze‐dried, and ground into a powder. For antioxidant assays, ground plant powder was extracted using 1% HCl, 90% aqueous methanol for 2 h under shaking. The 2,2‐diphenly‐1picryl‐hydrazyl‐hydrate (DPPH) and (ABTS) assays were used to determine the antioxidant capacity of each extract. Briefly, 10 μL extract was incubated with 10 μM DPPH for 10 min, after which the absorbance was measured spectrophotometrically at 517 nm. For the ABTS assay, 10 μL extract was incubated with 3.5 mM ABTS radical for 30 s. The absorbance at 734 nm was then measured spectrophotometrically. Results from both assays were compared to a trolox standard and calculated as 6‐hydroxy‐2,5,7,8‐tetramethylchroman‐2‐carboxylic acid (trolox) equivalents (TE; μmol/g dry weight). Results from the ABTS assay showed antioxidant activities of 189.5±26.7, 7462±286.3, 195.3±31.6, 85.03±19.7, and 484.1±29.3 TE (mean±SEM) for stems, flowers, leaves, roots, and seeds, respectively. In correlation, the DPPH assay revealed the same pattern of radical scavenging ability in the various tissues, with the flowers exhibiting the highest amount of antioxidant capacity. The stems, flowers, leaves, roots, and seeds exhibited 150.1±6.128, 910.8±31.62, 188.8±10.72, 118.2±13.27, and 486.9±17.81 TE (mean±SEM), respectively.The phenolic content of I. chinensis extracts was determined using the Folin‐Ciocalteu assay. 20 μL extract was incubated with 2N 10 μL extract reagent, ddH2O, and 12.56% sodium carbonate. After 30 min, the absorbance at 750 nm was measured spectrophotometrically. Results were compared to gallic acid as a standard, and the phenolic content was calculated as gallic acid equivalents (GAE; μmol/g dry weight). The phenolic content was 1127±42.6, 5046±151.0, 1206±49.3, 662.2±24.5, and 1270±70.5 GAE (mean±SEM).
Regulatory Role of Ceramide/mTOR Signaling in Exosome Secretion from Smooth Muscle Cells during Arterial Stiffening and Medial Calcification
Authors: Bhat Owais M. , Yuan Xinxu , Li Guangbi , Camus Sarah , Kukreja Rakesh C. , Gulbins Erich , Li Pin-Lan 
Published: April 2020

Increased secretion of exosomes from arterial smooth muscle cells (SMCs) has been shown to contribute to arterial medial calcification (AMC) associated with accumulation of calcium deposits in the arterial wall and osteoblastic differentiation of SMCs. In this study, we further explored the molecular mechanisms increasing exosome secretion during AMC, focusing on lysosomal‐ceramide/mTOR (mammalian target of rapamycin) signaling in the regulation of lysosome trafficking and exosome release. Smooth muscle‐specific lysosomal acid ceramidase (Ac) gene knock out (Asah1fl/fl/SMcre) mice were treated with a high dose of Vit D (500,000 IU/kg/day) for 4 days. By Alizarin Red S and Von Kossa staining, we observed significantly increased aortic medial calcification with augmented expression of osteogenic markers like osteopontin and RUNX2 in Asah1fl/fl/SMcre mice compared to their wild type littermates, which was markedly attenuated by pretreatment of these mice with Torin‐1, an mTOR inhibitor. Confocal microscopy showed Asah1fl/fl/SMcre mice had markedly increased colocalization of mTORC1 with Lamp‐1(lysosome marker), but decreased colocalization of VPS16 (a multivesicular bodies (MVBs) marker) with Lamp‐1, suggesting mTOR activation and reduced MVBs interaction with lysosomes. However, mTOR inhibition by Torin‐1 significantly reduced the colocalization of mTOR vs Lamp‐1 which inturn increased lysosome‐MVBs interaction. Torin‐1 was also found to reduce accumulation of CD63 and annexin 2 (AnX2) (exosome markers) in the coronary arterial wall of Asah1fl/fl/SMcre mice compared to wild type littermates. Using CASMCs, we observed that Ac gene deletion led to peripheral distribution of lysosomes and increased exosome secretion, which was blocked by Torin‐1. Functionally, Torin‐1 was found to significantly reduce arterial stiffening Asah1fl/fl/SMcre mice as shown by restoration from increased pulse wave velocity (PWV). Altogether, these results suggest that lysosomal ceramide‐mTOR signaling is critical for the control of lysosome trafficking, exosome secretion and arterial stiffening or osteogenesis.Support or Funding InformationSupported by NIH grants HL057244, HL075316 and DK120491
Development of Picomolar Affinity Antagonists and Long‐acting Agonists for the Adrenomedullin and CGRP Receptors Using Combinatorial Peptide Library and Structure‐guided Design Approaches
Authors: Pioszak Augen A. , Booe Jason M. , Warner Margaret L. 
Published: April 2020

Adrenomedullin (AM) and calcitonin gene‐related peptide (CGRP) have diverse functions in the cardiovascular, lymphatic, and nervous systems including regulation of vascular tone, cardioprotection, and pain transmission. These peptides signal through three heterodimeric receptors comprising the class B GPCR, the calcitonin receptor‐like receptor (CLR), and one of three receptor activity‐modifying proteins (RAMP1‐3) that determine ligand selectivity. CLR‐RAMP complexes are proven or promising drug targets for several diseases, however, the moderate binding affinities and short plasma half‐lives of AM and CGRP limit their utility as therapeutics. Here, we used synthetic peptide combinatorial library and rational structure‐guided design approaches to develop AM and CGRP variant peptides with dramatically enhanced receptor affinities. The binding of the variants to purified RAMP‐CLR extracellular domain (ECD) complexes was characterized and a high‐resolution crystal structure of an AM variant bound to RAMP2‐CLR ECD explained its enhanced affinity. Incorporation of the variants into different length AM and CGRP scaffolds yielded picomolar affinity antagonists or agonists that exhibited substantially prolonged cAMP signaling after ligand washout. These activities were demonstrated in a model cell line transiently expressing the receptors and in primary human cells or immortalized cell lines that naturally express the receptors. This work provides a suite of AM and CGRP variants that will be valuable pharmacological tools and may have promise as peptide therapeutics with long receptor residence times. In addition, the properties of the variants support a mechanism for RAMP function in which direct RAMP‐peptide contacts and allosteric modulation of CLR cooperate to determine ligand selectivity.Support or Funding InformationThis work was supported by grants NIH R01GM104251 and OCAST HR16‐005 (AAP) and AHA predoctoral fellowship 18PRE33990152 (JMB).
A Study on Morphometric Features of Trachea and Larynx
Authors: Alimohammadi Majid , Doroudi Majid , Ebrahimi Azarnoush , Zilabi Golsan 
Published: April 2020

Certain data such as weight and dimensions of structures are important in dosimetry for treating cancer patients using radiotherapy.The objective of this study is to determine the morphometric features of trachea and larynx and discuss the possible correlation between the height and weight of these structures with other parameters such as height, weight and age of patients. 29 samples were measured using measuring tape and digital scale. The results of measurements showed significant correlation between the age of individuals and the weight and length of trachea and larynx. However, no significant correlation was found between the length and weight of trachea or larynx with either gender, height or weight of individuals.
Orphan sensory receptors are expressed in hepatocytes
Authors: Shepard Blythe D. 
Published: April 2020

Sensory receptors, including olfactory receptors (ORs), taste receptors (TAS), and orphan G‐protein coupled receptors (GPRs) have recently been found in a variety of non‐sensory tissues where they have important physiological functions. As G protein‐coupled receptors (GPCRs), these proteins serve as chemosensors by interpreting and responding to chemical cues in their native environment (i.e. blood, urine) and can be activated by naturally occurring metabolites. We reasoned that the liver, the largest metabolic organ in the body, is primed to take advantage of some of these sensory receptors in order to regulate its internal environment. To identify the complete complement of sensory receptors expressed in murine liver, we performed an unbiased screen of whole liver using TaqMan qPCR array cards. This screen identified a total of 17 ORs, 10 TAS, 3 opsins, and 57 GPRs – with 7 of the GPRs expressed at levels equal to well‐characterized hepatic GPCRs such as glucagon receptor. In order to understand the functions of these sensory receptors, it is imperative to determine where they localize and their ligand profile. Due to the lack of reliable antibodies, we turned to RNAScope to localize mRNA transcript expression within the liver. Using this technique, we were able to confirm that murine Tas2r108, Olfr57, and Gpr137 are all expressed in hepatocytes, the metabolically active epithelial cells of the liver. With confirmation of expression, all three of these hepatic receptors were cloned from both male and female livers and expressed into heterologous cells for ligand screening. While Tas2r108 has known ligands – bitter tastants that have anti‐bacterial functions – both Olfr57 and Gpr137 remain orphan receptors. This is due, in part, to the inability to achieve functional expression of these receptors in vitro. Of note, Gpr137 has multiple splice variants and co‐expression of the canonical full‐length GPCR form along with a truncated variant increases surface expression of this 7‐transmembrane domain receptor. Ongoing studies are aimed at determining the role that Gpr137 plays in hepatic protein trafficking. Surface expression conditions were also achieved for Olfr57 with the assistance of a cleavable LUCY tag allowing for large‐scale ligand screening using a cAMP‐driven dual luciferase reporter assay. Using this platform, we determined that Olfr57 responds to terpenes including camphor and fenchol. These volatile organic compounds are known fungal metabolites, opening up the possibility that the liver is utilizing ORs to detect fungal growth. Efforts are currently underway to test this hypothesis and to establish the functional relevance for these chemosensors in hepatocytes.Support or Funding InformationNIDDK
Novel roles of Atrogin‐1 in cardiac disease, lipid metabolism and bone microstructure
Authors: Klüppel Michael , Suryadevara Vidyani , Willis Monte S. 
Published: April 2020

Ubiquitination plays a critical role in controlling protein turnover via proteasomal degradation, altering subcellular localization, and regulating protein activity. Functional role(s) for the Atrogin‐1 (MAFbx) ubiquitin ligase in regulating transcription factors have been found in skeletal muscle and the heart, but not in other organs. Within the myocyte, Atrogin‐1 localizes to the sarcomere and nucleus. We have previously reported that Atrogin‐1 is a critical regulator of pathologic and physiologic cardiac hypertrophy in vivo, involving specific interaction and ubiquitination of transcription factors central the signaling pathways driving these hypertrophic responses, including FOXO1/3. Recent studies have linked cardiomyocyte Atrogin‐1 to the regulation of the extracellular matrix. With evidence that cardiomyocyte Atrogin‐1 can affect cells other than myocytes, we analyzed Atrogin‐1−/− mice for their effects on metabolism and bone. DEXA whole body analysis of Atrogin‐1−/− mice revealed significantly decreased fat mass (15.6% vs. 35.8% fat, p<0.05) with a corresponding increase in lean mass (84.4% vs. 64.2%, p<0.05) at the age of 11–15 months, but not at 5 months, when compared to strain‐ and age‐matched wildtype (WT) control mice. Since the Atrogin‐1−/− mice weighed significantly less than WT mice (29±3 vs. 40±2 g, p<0.05), the net fat loss was 4.6 g (vs. 14.3 g in age‐matched WT) and the lean body mass identical between groups (25.7 vs. 24.6 g). Together these findings suggest Atrogin‐1’s role in regulating fat metabolism. Analysis of bone microarchitecture and mechanical properties of adult Atrogin‐1−/− mice was undertaken at 17–21 weeks of age, a time point where no phenotype has been previously observed in skeletal muscle or heart. Femurs were dissected and subsequently assessed by μCT (SkyScan 1172) for bone microarchitecture in the distal femur metaphysis (1 mm) and mid‐diaphysis region and challenged by a three‐point bending test to assess their material and structural properties. No differences in cortical or trabecular bone microarchitecture were identified and nor were any changes in mechanical properties. Together, these findings illustrate novel biological roles of Atrogin‐1 in regulating systemic fat metabolism, possibly involving crosstalk between skeletal muscle/heart and systemic fat metabolism. Our analysis provides an essential framework for the potential therapeutic targeting of ubiquitin ligases like Atrogin‐1 in the context of striated muscle and metabolic disease.Support or Funding InformationSupport: Lilly Foundation/Indiana Center for Musculoskeletal Health/ IU School of Medicine, Physician Scientist Initiative, Scientific Research Initiative
Chemerin Increases Contractility of the Uterine Artery and Leads to Placentomegalia
Authors: Justina Vanessa Dela , Priviero Fernanda , Lima Victor Vitorino , Giachini Fernanda Regina , Webb R. Clinton 
Published: April 2020

Chemerin is an adipokine released from several tissues (mostly adipocytes but also placenta, ovaries and liver) which participates in the regulation of adipocyte development, inflammatory processes, and metabolic functions. Chemerin is gaining attention in gestation and it has been reported to be augmented in preeclampsia. This adipokine plays a physiological and protective role during early pregnancy; however, late complications in pregnancy may be observed when chemerin levels remain augmented. During pregnancy, maternal uterine artery (UA) blood flow is critical to maintaining the intrauterine environment, permitting normal placental function, and supporting fetal growth. Chemerin has been shown to contract many vascular beds, however, its role on the UA remains to be elucidated. This study tests the hypothesis that chemerin leads to increased contraction of UA contributing to the development of preeclampsia‐like symptoms and leading to gestational disorders. Pregnant (P) and non‐pregnant (NP) Sprague Dawley rats (200–250g) were injected daily (i.p.) with vehicle (saline) or chemerin (16 μg/rat;). On gestational day 20, or corresponding day for NP rats, animals were euthanized and placentas and fetuses weighted. UA was removed, cleaned and mounted in wire myograph. Concentration response‐curves to acetylcholine (ACh) and seretonin [5‐HT (1 nM to 100 μM); in the presence or absence of ERK1/2 inhibitor, PD98059] were performed in UA. Treatment with chemerin decreases the relaxation to ACh in both P [Emax 69.77 ± 4.90 vs. 95.50 ± 3.10] and NP [Emax 74.56 ± 4.52 vs. 96.04 ± 5.75] animals compared to respective controls. Increased sensitivity to 5‐HT was observed in the UAs of chemerin‐treated P [pD2 7.05 ± 0.05 vs. 6.60 ± 0.16] and NP [pD2 6.57 ± 0.07 vs. 6.14 ± 0.14] rats compared to vehicle rats, which was abolished by incubation with PD98059. Further, chemerin treatment on P animals resulted in increased placental weight [(g) 0.50 ± 0.016 vs. 0.43 ± 0.004] with no changes in fetal weight. We conclude that chemerin treatment impairs UA contractility and relaxation leading to placental disorder. These changes in UA function may be a predictive for elevate blood pressure in preeclampsia.Support or Funding InformationNIH 13604