Testosterone regulates
            CYP2J19
            -linked carotenoid signal expression in male red-backed fairywrens (
            Malurus melanocephalus
            )

Carotenoid pigments produce most red, orange and yellow colours in vertebrates. This coloration can serve as an honest signal of quality that mediates social and mating interactions, but our understanding of the underlying mechanisms that control carotenoid signal production, including how different physiological pathways interact to shape and maintain these signals, remains incomplete. We investigated the role of testosterone in mediating gene expression associated with a red plumage sexual signal in red-backed fairywrens ( Malurus melanocephalus ). In this species, males within a single population can flexibly produce either red/black nuptial plumage or female-like brown plumage. Combining correlational analyses with a field-based testosterone implant experiment and quantitative polymerase chain reaction, we show that testosterone mediates expression of carotenoid-based plumage in part by regulating expression of CYP2J19 , a ketolase gene associated with ketocarotenoid metabolism and pigmentation in birds. This is, to our knowledge, the first time that hormonal regulation of a specific genetic locus has been linked to carotenoid production in a natural context, revealing how endocrine mechanisms produce sexual signals that shape reproductive success.

Download Full-text

Cyclo-oxygenase 2, a putative mediator of vessel remodeling, is expressed in the brain AVM vessels and associates with inflammation

Acta Neurochirurgica ◽

10.1007/s00701-021-04895-z ◽

2021 ◽

Author(s):

Sara Keränen ◽

Santeri Suutarinen ◽

Rahul Mallick ◽

Johanna P. Laakkonen ◽

Diana Guo ◽

...

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Rank Correlation ◽

Inflammatory Cells ◽

Vessel Remodeling ◽

Brain Avm ◽

Inflammatory Drugs ◽

Polymerase Chain ◽

The Brain ◽

Cox2 Expression

Abstract Background Brain arteriovenous malformations (bAVM) may rupture causing disability or death. BAVM vessels are characterized by abnormally high flow that in general triggers expansive vessel remodeling mediated by cyclo-oxygenase-2 (COX2), the target of non-steroidal anti-inflammatory drugs. We investigated whether COX2 is expressed in bAVMs and whether it associates with inflammation and haemorrhage in these lesions. Methods Tissue was obtained from surgery of 139 bAVMs and 21 normal Circle of Willis samples. The samples were studied with immunohistochemistry and real-time quantitative polymerase chain reaction (RT-PCR). Clinical data was collected from patient records. Results COX2 expression was found in 78% (109/139) of the bAVMs and localized to the vessels’ lumen or medial layer in 70% (95/135) of the bAVMs. Receptors for prostaglandin E2, a COX2-derived mediator of vascular remodeling, were found in the endothelial and smooth muscle cells and perivascular inflammatory cells of bAVMs. COX2 was expressed by infiltrating inflammatory cells and correlated with the extent of inflammation (r = .231, p = .007, Spearman rank correlation). COX2 expression did not associate with haemorrhage. Conclusion COX2 is induced in bAVMs, and possibly participates in the regulation of vessel wall remodelling and ongoing inflammation. Role of COX2 signalling in the pathobiology and clinical course of bAVMs merits further studies.

Download Full-text

Type III Collagen is Required for Adipogenesis and Actin Stress Fibre Formation in 3T3-L1 Preadipocytes

Biomolecules ◽

10.3390/biom11020156 ◽

2021 ◽

Vol 11 (2) ◽

pp. 156

Author(s):

Mohammad Al Hasan ◽

Patricia E. Martin ◽

Xinhua Shu ◽

Steven Patterson ◽

Chris Bartholomew

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Type Iii Collagen ◽

Type Iii ◽

Actin Stress Fibre ◽

Matrix Gene ◽

Gene Transcripts ◽

Polymerase Chain ◽

Oil Red O Staining ◽

Oil Red O

GPR56 is required for the adipogenesis of preadipocytes, and the role of one of its ligands, type III collagen (ColIII), was investigated here. ColIII expression was examined by reverse transcription quantitative polymerase chain reaction, immunoblotting and immunostaining, and its function investigated by knockdown and genome editing in 3T3-L1 cells. Adipogenesis was assessed by oil red O staining of neutral cell lipids and production of established marker and regulator proteins. siRNA-mediated knockdown significantly reduced Col3a1 transcripts, ColIII protein and lipid accumulation in 3T3-L1 differentiating cells. Col3a1−/− 3T3-L1 genome-edited cell lines abolished adipogenesis, demonstrated by a dramatic reduction in adipogenic moderators: Pparγ2 (88%) and C/ebpα (96%) as well as markers aP2 (93%) and oil red O staining (80%). Col3a1−/− 3T3-L1 cells displayed reduced cell adhesion, sustained active β-catenin and deregulation of fibronectin (Fn) and collagen (Col4a1, Col6a1) extracellular matrix gene transcripts. Col3a1−/− 3T3-L1 cells also had dramatically reduced actin stress fibres. We conclude that ColIII is required for 3T3-L1 preadipocyte adipogenesis as well as the formation of actin stress fibres. The phenotype of Col3a1−/− 3T3-L1 cells is very similar to that of Gpr56−/− 3T3-L1 cells, suggesting a functional relationship between ColIII and Gpr56 in preadipocytes.

Download Full-text

Decreased Expression of GPER1 Gene and Protein in Goiter

International Journal of Endocrinology ◽

10.1155/2015/869431 ◽

2015 ◽

Vol 2015 ◽

pp. 1-5 ◽

Cited By ~ 2

Author(s):

Raquel Weber ◽

Ana Paula Santin Bertoni ◽

Laura Walter Bessestil ◽

Ilma Simoni Brum ◽

Tania Weber Furlanetto

Keyword(s):

Estrogen Receptor ◽

Quantitative Polymerase Chain Reaction ◽

Thyroid Tissue ◽

Chain Reaction ◽

Normal Thyroid ◽

Protein Levels ◽

Protein Expressions ◽

Polymerase Chain ◽

G Protein Coupled

Goiter is more common in women, suggesting that estrogen could be involved in its physiopathology. The presence of classical estrogen receptors (ERαand ERβ) has been described in thyroid tissue, suggesting a direct effect of estrogen on the gland. A nonclassic estrogen receptor, the G-protein-coupled estrogen receptor (GPER1), has been described recently in several tissues. However, in goiter, the presence of this receptor has not been studied yet. We investigated GPER1 gene and protein expressions in normal thyroid and goiter using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot, respectively. In normal thyroid (n=16) and goiter (n=19), GPER1 gene was expressed in all samples, while GPER1 protein was expressed in all samples of normal thyroid (n=15) but in only 72% of goiter samples (n=13). When comparing GPER1 gene and protein levels in both conditions, gene expression and protein levels were higher in normal thyroid than in goiter, suggesting a role of this receptor in this condition. Further studies are needed to elucidate the role of GPER1 in normal thyroid and goiter.

Download Full-text

C/EBPα directs monocytic commitment of primary myeloid progenitors

Blood ◽

10.1182/blood-2005-12-008763 ◽

2006 ◽

Vol 108 (4) ◽

pp. 1223-1229 ◽

Cited By ~ 65

Author(s):

Dehua Wang ◽

Jenice D'Costa ◽

Curt I. Civin ◽

Alan D. Friedman

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Fold Increase ◽

Pcr Assay ◽

Gm Csf ◽

Specific Effects ◽

Proliferation And Apoptosis ◽

Polymerase Chain ◽

Lineage Markers ◽

Transcriptional Induction

Abstract C/EBPα is required for generation of granulocyte-monocyte progenitors, but the subsequent role of C/EBPα in myeloid lineage commitment remains uncertain. We transduced murine marrow cells with C/EBPα-estradiol receptor (ER) or empty vector and subjected these to lineage depletion just prior to culture in estradiol with myeloid cytokines. This protocol limits biases due to lineage-specific effects on developmental kinetics, proliferation, and apoptosis. Also, lowering the dose of estradiol reduced activated C/EBPα-ER to near the physiologic range. C/EBPα-ER increased Mac1+/Gr1–/MPO–/low monocytes 1.9-fold while reducing Mac1+/Gr1+/MPOhi granulocytes 2.5-fold at 48 hours, even in 0.01 μM estradiol. This pattern was confirmed morphologically and by quantitative polymerase chain reaction (PCR) assay of lineage markers. To directly assess effects on immature progenitors, transduced cells were cultured for 1 day with and then in methylcellulose without estradiol. A 2-fold increase in monocytic compared with granulocytic colonies was observed in IL-3/IL-6/SCF or GM-CSF, but not G-CSF, even in 0.01 μM estradiol. C/EBPα-ER induced PU.1 mRNA, and PU.1-ER stimulated monocytic development, suggesting that transcriptional induction of PU.1 by C/EBPα contributes to monopoiesis. A C/EBPα variant incapable of zippering with c-Jun did not induce monopoiesis, and a variant unable to bind NF-κB p50 stimulated granulopoiesis, suggesting their cooperation with C/EBPα during monocytic commitment.

Download Full-text

Sinonasal Surfactant Protein A1, A2, and D Gene Expression in Cystic Fibrosis: A Preliminary Report

Otolaryngology ◽

10.1016/j.otohns.2007.01.025 ◽

2007 ◽

Vol 137 (1) ◽

pp. 34-38 ◽

Cited By ~ 18

Author(s):

Bradford A. Woodworth ◽

Rachel Wood ◽

John E. Baatz ◽

Rodney J. Schlosser

Keyword(s):

Gene Expression ◽

Cystic Fibrosis ◽

Nasal Polyposis ◽

Quantitative Polymerase Chain Reaction ◽

Surfactant Protein ◽

Fungal Rhinosinusitis ◽

Allergic Fungal Rhinosinusitis ◽

Novel Treatment ◽

Polymerase Chain

OBJECTIVE: To measure alterations in SPA1, A2, and D gene expression in various forms of inflammatory chronic rhinosinusitis (CRS). STUDY DESIGN AND SETTING: Sinus mucosal biopsies were performed in patients with allergic fungal rhinosinusitis (AFS), CRS with nasal polyposis, cystic fibrosis (CF), and controls. SP mRNA was measured with quantitative polymerase chain reaction. RESULTS: Patients with CF (n = 4) showed significantly increased SPA1 (82-fold), SPA2 (100-fold), and SPD (47-fold) mRNA ( P < 0.05) when compared with controls (n = 5). Patients with CRS with nasal polyposis (n = 5) also demonstrated elevated SPA1 (27-fold), SPA2 (13-fold), and SPD (13-fold). Patients with AFS (n = 7) had increased SPA1 (5-fold), SPA2 (9-fold), and SPD (17-fold), but were not statistically significant. CONCLUSION: SPA1, A2, and D are upregulated in various forms of CRS, but are significantly elevated in cystic fibrosis CRS. SIGNIFICANCE: Understanding the role of SPs in CRS will help develop novel treatment approaches for sinonasal pathoses.

Download Full-text

Activation of C-Type Lectin Receptor and (RIG)-I-Like Receptors Contributes to Proinflammatory Response in Middle East Respiratory Syndrome Coronavirus-Infected Macrophages

The Journal of Infectious Diseases ◽

10.1093/infdis/jiz483 ◽

2019 ◽

Cited By ~ 2

Author(s):

Xiaoyu Zhao ◽

Hin Chu ◽

Bosco Ho-Yin Wong ◽

Man Chun Chiu ◽

Dong Wang ◽

...

Keyword(s):

Middle East ◽

Quantitative Polymerase Chain Reaction ◽

Severe Disease ◽

Human Infection ◽

Middle East Respiratory Syndrome ◽

Proinflammatory Response ◽

Pharmacological Inhibition ◽

Lectin Receptor ◽

Polymerase Chain

Abstract Background Human infection with Middle East respiratory syndrome coronavirus (MERS-CoV) poses an ongoing threat to public health worldwide. The studies of MERS patients with severe disease and experimentally infected animals showed that robust viral replication and intensive proinflammatory response in lung tissues contribute to high pathogenicity of MERS-CoV. We sought to identify pattern recognition receptor (PRR) signaling pathway(s) that mediates the inflammatory cascade in human macrophages upon MERS-CoV infection. Methods The potential signaling pathways were manipulated individually by pharmacological inhibition, small interfering ribonucleic acid (siRNA) depletion, and antibody blocking. The MERS-CoV-induced proinflammatory response was evaluated by measuring the expression levels of key cytokines and/or chemokines. Reverse transcription-quantitative polymerase chain reaction assay, flow cytometry analysis, and Western blotting were applied to evaluate the activation of related PRRs and engagement of adaptors. Results MERS-CoV replication significantly upregulated C-type lectin receptor (CLR) macrophage-inducible Ca2+-dependent lectin receptor (Mincle). The role of Mincle for MERS-CoV-triggered cytokine/chemokine induction was established based on the results of antibody blockage, siRNA depletion of Mincle and its adaptor spleen tyrosine kinase (Syk), and Syk pharmacological inhibition. The cytokine and/or chemokine induction was significantly attenuated by siRNA depletion of retinoic acid-inducible-I-like receptors (RLR) or adaptor, indicating that RLR signaling also contributed to MERS-CoV-induced proinflammatory response. Conclusions The CLR and RLR pathways are activated and contribute to the proinflammatory response in MERS-CoV-infected macrophages.

Download Full-text

Occurrence of a 16SrIV Group Phytoplasma not Previously Associated with Palm Species in Yucatan, Mexico

Plant Disease ◽

10.1094/pdis-02-10-0150 ◽

2011 ◽

Vol 95 (3) ◽

pp. 256-262 ◽

Cited By ~ 24

Author(s):

Roberto Vázquez-Euán ◽

Nigel Harrison ◽

María Narvaez ◽

Carlos Oropeza

Keyword(s):

Cocos Nucifera ◽

Restriction Enzymes ◽

Nested Polymerase Chain Reaction ◽

Palm Trees ◽

Palm Species ◽

Leaf Yellowing ◽

Sabal Mexicana ◽

Polymerase Chain ◽

First Time

The occurrence of 16SrIV group phytoplasmas in palm species Sabal mexicana and Pseudophoenix sargentii is reported here for the first time. Palm trees showed leaf decay and leaf yellowing syndromes, respectively. An amplification product (1.4 kb) was obtained in symptomatic S. mexicana (18 of 21) and symptomatic P. sargentii (1 of 1) palm trees sampled in different locations in Yucatan State, Mexico; five of the positive S. mexicana and the positive P. sargentii trees died. The identity of the phytoplasmas from these species was determined by restriction fragment length polymorphism profiling with restriction enzymes AluI and HinfI, showing there could be two phytoplasma strains of the 16SrIV group. In one S. mexicana palm, the profile was the same as observed with these enzymes for phytoplasmas of 16SrIV-A subgroup, previously associated with Cocos nucifera palm trees and, in the rest of the trees, including the P. sargentii palm, the profile was for phytoplasmas of the 16SrIV-D subgroup. These identities were supported by analyses of the amplicons obtained by nested polymerase chain reaction by nucleotide-nucleotide BLAST analysis. Geographical distribution of the association S. mexicana/16SrIV group phytoplasmas was found widely dispersed in Yucatan State. A potential role of S. mexicana palm trees as a permanent source of phytoplasma inoculum is suggested. In addition to P. sargentii, other palm species (Thrinax radiata and C. nucifera) coexisting with S. mexicana trees were also sampled and analyzed.

Download Full-text

Possible Role of Trichothecene Mycotoxins in Virulence of Fusarium graminearum on Maize

Plant Disease ◽

10.1094/pdis.1999.83.10.954 ◽

1999 ◽

Vol 83 (10) ◽

pp. 954-960 ◽

Cited By ~ 130

Author(s):

L. J. Harris ◽

A. E. Desjardins ◽

R. D. Plattner ◽

P. Nicholson ◽

G. Butler ◽

...

Keyword(s):

Fusarium Graminearum ◽

Fungal Biomass ◽

Biosynthetic Pathway ◽

Quantitative Polymerase Chain Reaction ◽

Field Trials ◽

Ear Rot ◽

Inoculation Method ◽

Trichodiene Synthase ◽

Polymerase Chain

Trichothecene-producing and -nonproducing Fusarium graminearum strains were tested for their ability to cause Gibberella ear rot in field trials at two locations—Ottawa, Ontario, and Peoria, Illinois—in 1996. Maize ears were inoculated with wild-type or transgenic F. graminearum strains in which the trichothecene biosynthetic pathway had been disabled by the specific disruption of the trichodiene synthase gene and with a derivative revertant strain in which trichothecene production had been restored through recombination. A silk channel inoculation method was employed at both locations. In addition, a kernel puncture inoculation method was used at the Ontario location. Harvested maize ears were analyzed for visual disease severity, grain yield, deoxynivalenol (DON) concentration, and fungal biomass by quantitative polymerase chain reaction (PCR) and/or ergosterol quantitation. There was a significant correlation (r= 0.86) between data obtained from the two different methods of quantifying fungal biomass. The trichothecene-nonproducing strains were still pathogenic but appeared less virulent on maize than the trichothecene-producing progenitor and revertant strains, as assayed by most parameters. This suggests that the trichothecenes may act as virulence factors to enhance the spread of F. graminearum on maize.

Download Full-text

β-Adrenergic receptor stimulation inhibits proarrhythmic alternans in postinfarction border zone cardiomyocytes: a computational analysis

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00094.2017 ◽

2017 ◽

Vol 313 (2) ◽

pp. H338-H353 ◽

Cited By ~ 17

Author(s):

Jakub Tomek ◽

Blanca Rodriguez ◽

Gil Bub ◽

Jordi Heijman

Keyword(s):

Adrenergic Receptor ◽

Border Zone ◽

Adrenergic Stimulation ◽

Cell Coupling ◽

Postmyocardial Infarction ◽

Model Cell ◽

Cycle Lengths ◽

Underlying Mechanisms ◽

First Time

The border zone (BZ) of the viable myocardium adjacent to an infarct undergoes extensive autonomic and electrical remodeling and is prone to repolarization alternans-induced cardiac arrhythmias. BZ remodeling processes may promote or inhibit Ca2+ and/or repolarization alternans and may differentially affect ventricular arrhythmogenesis. Here, we used a detailed computational model of the canine ventricular cardiomyocyte to study the determinants of alternans in the BZ and their regulation by β-adrenergic receptor (β-AR) stimulation. The BZ model developed Ca2+ transient alternans at slower pacing cycle lengths than the control model, suggesting that the BZ may promote spatially heterogeneous alternans formation in an infarcted heart. β-AR stimulation abolished alternans. By evaluating all combinations of downstream β-AR stimulation targets, we identified both direct (via ryanodine receptor channels) and indirect [via sarcoplasmic reticulum (SR) Ca2+ load] modulation of SR Ca2+ release as critical determinants of Ca2+ transient alternans. These findings were confirmed in a human ventricular cardiomyocyte model. Cell-to-cell coupling indirectly modulated the likelihood of alternans by affecting the action potential upstroke, reducing the trigger for SR Ca2+ release in one-dimensional strand simulations. However, β-AR stimulation inhibited alternans in both single and multicellular simulations. Taken together, these data highlight a potential antiarrhythmic role of sympathetic hyperinnervation in the BZ by reducing the likelihood of alternans and provide new insights into the underlying mechanisms controlling Ca2+ transient and repolarization alternans. NEW & NOTEWORTHY We integrated, for the first time, postmyocardial infarction electrical and autonomic remodeling in a detailed, validated computer model of β-adrenergic stimulation in ventricular cardiomyocytes. Here, we show that β-adrenergic stimulation inhibits alternans and provide novel insights into underlying mechanisms, adding to a recent controversy about pro-/antiarrhythmic effects of postmyocardial infarction hyperinnervation. Listen to this article’s corresponding podcast at http://ajpheart.podbean.com/e/%CE%B2-ar-stimulation-and-alternans-in-border-zone-cardiomyocytes/ .

Download Full-text

Integrative Analysis of CircRNA Expression and Associated ceRNA Network in Graves’ Disease

10.21203/rs.3.rs-191564/v1 ◽

2021 ◽

Author(s):

Zhengrong Jiang ◽

Linghong Huang ◽

Lijun Chen ◽

Jingxiong Zhou ◽

Xuefeng Bai ◽

...

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Family Members ◽

Circular Rnas ◽

Newly Diagnosed ◽

Autoimmune Thyroid ◽

Competing Endogenous Rnas ◽

Healthy Control ◽

Polymerase Chain ◽

Comprehensive Study

Abstract ObjectiveGraves’ disease (GD) is the most common subtype of autoimmune thyroid disease which have the involvement of circular RNAs (circRNAs) in the pathogenesis. However, it is largely unknown about the role of circRNAs in GD. In present study, we performed a comprehensive study of the circRNAs in GD by bioinformatics analyses.MethodsCircRNAs were detected in plasma of 3 newly diagnosed GD patients and 3 healthy controls. We selected 2 up-regulated and 1 down-regulated circRNAs with different expression in GD for validation by transcriptase-quantitative polymerase chain reaction in both GD and healthy control subjects. The GTRD base was used for predicting the transcript factors of the different expression of circRNAs. Then the competing endogenous RNAs(ceRNAs) network was assembled and the analysis of the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes were performed to predict the functions of different expression of circRNAs.ResultsA total of 366 different expression of circRNAs were detected in GD. The exonic circRNA hsa_circ_0090364 was validated as an up-regulated molecule in GD. 82 transcript factors in promoter region for hsa_circ_0090364 were predicted. The ceRNAs network revealed that the miR-378 family members acted as the target microRNAs of hsa_circ_0090364 and its downstream genes, and also unveiled the involvement of hsa_circ_0090364 in pathogenesis of GD.ConclusionsHsa_circ_0090364 might play a role of ceRNA through sponging up miR-378 family members, which demonstrated that circRNAs had an effect on regulation in GD.

Download Full-text