Journal: Archives of Gynecology and Obstetrics

Article Title: Glycosylated fibronectin as a first trimester marker for gestational diabetes

doi: 10.1007/s00404-020-05670-8

Figure Lengend Snippet: a Glycosylated fibronectin concentrations in BMI subgroups of 20–25 and 30–35 kg/m 2 in the study and the control group. b Effect of smoking on concentrations of glycosylated fibronectin in the control and GDM groups. c Multiple of median (MoM) values for fibronectin and glycosylated fibronectin between control and GDM groups. d Multiple of median (MoM) values for adiponectin and glycosylated adiponectin between control and GDM groups

Article Snippet: Calibrators for both assays were made from recombinant human adiponectin (1065-AF, R&D Systems, Abingdon, UK).

Techniques: Control

Journal: Diabetes

Article Title: Adiponectin Reduces Plasma Triglyceride by Increasing VLDL Triglyceride Catabolism

doi: 10.2337/db07-0435

Figure Lengend Snippet: Elevated adiponectin has no effect on HNF4α protein levels in mouse liver. Three days after viral vector injection, liver tissues were collected in the fasting state after perfusion. Total liver protein was prepared, and HNF4α and adiponectin were analyzed by Western blotting.

Article Snippet: Recombinant human adiponectin and anti-mouse adiponectin antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Plasmid Preparation, Injection, Western Blot

Journal: PLoS ONE

Article Title: A Saccharomyces cerevisiae Assay System to Investigate Ligand/AdipoR1 Interactions That Lead to Cellular Signaling

doi: 10.1371/journal.pone.0065454

Figure Lengend Snippet: (A) The components of the AdipoR1 signaling pathway of mammalian cells are shown in black font and their S. cerevisiae homologs, if known are shown in red font. In mammalian cells, adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif 1 (APPL1) interacts directly with AdipoR1. Interaction of adiponectin (ADPN) with AdipoR1 stimulates AdipoR1-APPL1 interaction. This results in release of Ca 2+ from the ER to the cytosol and also increases export of LKB1 kinase from the nucleus. Influx of extracellular Ca 2+ to the cytosol is also stimulated by adiponectin, although the mechanism by which this occurs remains to be clarified. Increase in cytosolic Ca 2+ concentration activates Ca 2+ /calmodulin-dependent protein kinase kinase (CaMKK) which in turn activates AMP activated protein kinase (AMPK) by phosphorylating its α subunit. However, phosphorylation of AMPK α subunit by the cytosol-localized kinase LKB1 is the major pathway for activation of AMPK. Adiponectin-AdipoR1 interaction also increases cellular ceramidase activity which in turn leads to phosphorylation of AMPKα subunit. The details of this pathway are not clear yet. Activation of AMPK is required for many of the anti-diabetic and anti-atherosclerotic effects of adiponectin. In vascular endothelial cells, interaction of AdipoR1 with adiponectin activates protein kinase A (PKA) which has the effect of lowering accumulation of reactive oxygen species (ROS) and thereby reducing inflammation. (B) Subunit structure of S. cerevisiae AMPK (ScAMPK). Like mammalian AMPK, ScAMPK is a trimer composed of an α, β, and γ subunit. The genes encoding the β subunit isoforms as well as the sole α and γ subunits are indicated. (C) Components of the IZH2 -mediated signaling pathways in S. cerevisiae are shown in red font. Interaction of IZH2 (homolog of AdipoRs) with osmotin (OSM) activates PKA via a RAS2-cAMP pathway. Overexpression of IZH2 , overexpression of AdipoR1 , treatment of S. cerevisiae cells expressing AdipoR1 with adiponectin and treatment of S. cerevisiae cells expressing various levels of IZH2 with thaumatin (THN, a homolog of osmotin) has been shown to activate PKA by increasing cellular ceramidase activity. Activation of PKA leads to decreased transcription from a stress responsive promoter element ( STRE ) and increased cellular ROS content. Activated PKA promotes export of ScAMPK from the nucleus which leads to decreased transcription from the ferroxidase (FET3 ) promoter. Activated PKA also represses FET3 transcription via the stress-responsive transcription factors MSN2/4. Mutational analyses show that genes encoding the APPL1-lke protein Sip3, the LKB1-like protein Sak1, the ScAMPK β subunit Sip1 and the ScAMPK γ subunit Snf4 are components of the pathway leading from IZH2 (or AdipoR1 ) to FET3 repression in S. cerevisiae .

Article Snippet: Unless specified otherwise, the pure recombinant Escherichia coli expressed N-terminal (His) 6 -tagged full length human adiponectin used in these studies was the gift of Dr. A. Sharkhuu (KAUST, Saudi Arabia) or Dr.

Techniques: Binding Assay, Concentration Assay, Phospho-proteomics, Activation Assay, Activity Assay, Protein-Protein interactions, Over Expression, Expressing

Journal: PLoS ONE

Article Title: A Saccharomyces cerevisiae Assay System to Investigate Ligand/AdipoR1 Interactions That Lead to Cellular Signaling

doi: 10.1371/journal.pone.0065454

Figure Lengend Snippet: Cells of strain BY4741carrying pESC-URA-CLuc-AdipoR1-APPL1-NLuc were grown for 16 h at 30°C in selective minimal medium at the indicated galactose concentrations, treated for 4 h at 30°C with the indicated test compounds and then assayed for Luc activity. (A) Imaging of Luc activity. (B) Quantitative measurement of Luc activity as a function of osmotin concentration. A representative image of relative Luc activity at the different osmotin concentrations is shown for each galactose concentration. Data represent the means ± SD from three experiments with triplicate samples. For each galactose concentration, significant differences by a Student’s t-test between osmotin treated samples and untreated control are indicated by asterisks. Symbols: PBS, 1/8 X PBS; f-ADPN, bacterially expressed full length adiponectin; BSA, bovine serum albumin, OSM, osmotin; g-ADPN, bacterially expressed globular adiponectin; **, p <0.01; ***, p <0.001.

Article Snippet: Unless specified otherwise, the pure recombinant Escherichia coli expressed N-terminal (His) 6 -tagged full length human adiponectin used in these studies was the gift of Dr. A. Sharkhuu (KAUST, Saudi Arabia) or Dr.

Techniques: Activity Assay, Imaging, Concentration Assay, Control

Journal: PLoS ONE

Article Title: A Saccharomyces cerevisiae Assay System to Investigate Ligand/AdipoR1 Interactions That Lead to Cellular Signaling

doi: 10.1371/journal.pone.0065454

Figure Lengend Snippet: (A) Cell lysates (100 µg protein) of strain BWG1-7a and an isogenic Δsnf1::Kan_MX line were fractionated by 10% SDS-PAGE. Shown are blots probed with anti-phospho-AMPK(Thr-172) antibody. The expected size of Snf1p is 72 kDa. (B, C) Cells (about 10 8 /mL) of strain BWG1-7a Δizh2::Kan_MX transformed with p426GPD (Vec), p426GPD- AdipoR1 (pAdipoR1) or p426GPD- AdipoR2 (pAdipoR2) were treated with indicated osmotin and adiponectin concentrations for 30 min at 30°C in YPD. Aliquots were withdrawn for viable counts determination before the cell lysates were prepared for analysis by10% SDS-PAGE. Shown in the top panels are blots probed first with phospho-AMPK(Thr-172) antibody, then stripped and probed with actin antibody (100 µg total protein per lane). Shown in the middle panels are relative band intensities in the depicted gels. ‘Relative band intensity’ was defined as the ratio of the intensity of phospho-Snf1p signal to actin signal for each lane when the value of this ratio for the corresponding untreated Vec sample was arbitrarily assigned the value 1.0. Shown in the bottom panels are viable counts in each sample at the end of the treatments. The experiments were performed twice with comparable results and the results of one experiment are shown.

Article Snippet: Unless specified otherwise, the pure recombinant Escherichia coli expressed N-terminal (His) 6 -tagged full length human adiponectin used in these studies was the gift of Dr. A. Sharkhuu (KAUST, Saudi Arabia) or Dr.

Techniques: SDS Page, Transformation Assay

Journal: Annals of Dermatology

Article Title: Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis

doi: 10.5021/ad.2019.31.2.186

Figure Lengend Snippet: Schedule to develop in vitro cultured human epidermal equivalents (HEEs) model mimicking atopic dermatitis (AD). During the preparation of reconstructed three-dimensional epidermis, the cocktail containing Poly(I:C) and inflammatory cytokines was treated on the 11th day after air-lift to generate AD-HEEs. Adiponectin was co-treated with the cocktail at the same time to investigate the effect of adiponectin on AD conditioned epidermis. And then the samples were harvested for analysis 3 days later. Tx.: treatment, TLR: Toll-like receptor 3, TNF-α: tumor necrosis factor-alpha, IL: interleukin.

Article Snippet: Recombinant human full-length adiponectin protein was obtained from Biobud (Seongnam, Korea).

Techniques: In Vitro, Cell Culture

Journal: Annals of Dermatology

Article Title: Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis

doi: 10.5021/ad.2019.31.2.186

Figure Lengend Snippet: Immunohistochemical staining of normal skin and atopic dermatitis lesional skin for adiponectin receptors. Protein expression of (A) adiponectin receptor-1 (AdipoR1) and (B) -2 (AdipoR2) in human epidermis were diminished in 3 atopic dermatitis patients, compared to control epidermis from 3 normal young adults (n=3 in each group). Bar=100 µm.

Article Snippet: Recombinant human full-length adiponectin protein was obtained from Biobud (Seongnam, Korea).

Techniques: Immunohistochemical staining, Staining, Expressing, Control

Journal: Annals of Dermatology

Article Title: Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis

doi: 10.5021/ad.2019.31.2.186

Figure Lengend Snippet: Effects of adiponectin on inflammatory markers in three-dimensional skin atopic dermatitis (AD) model. The quantitative real-time reverse transcriptase polymerase chain reaction analysis showed the changes in the expression of major inflammatory markers by adiponectin treatment. (A) carbonic anhydrase II (CAII), (B) neuron-specific NEL-like protein 2 (NELL2), (C) thymic stromal lymphopoietin (TSLP) were evaluated as the specific inflammatory markers for atopic dermatitis. Others, (D) human beta-defensin 2 (hBD2), (E) interleukin (IL)-8, and (F) tumor necrosis factor-alpha (TNF-α) were assessed as the primary inflammatory markers. * p <0.05, ** p <0.01.

Article Snippet: Recombinant human full-length adiponectin protein was obtained from Biobud (Seongnam, Korea).

Techniques: Reverse Transcription, Polymerase Chain Reaction, Expressing

Journal: Annals of Dermatology

Article Title: Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis

doi: 10.5021/ad.2019.31.2.186

Figure Lengend Snippet: Representative microscopic finding of (A) normal untreated control, (B) control for reconstructed epidermal atopic dermatitis (AD) model (only the inflammatory cocktail treated), and (C) the adiponectin treated sample (the inflammatory cocktail and adiponectin co-treated). HEEs: human epidermal equivalents, Tx.: treatment.

Article Snippet: Recombinant human full-length adiponectin protein was obtained from Biobud (Seongnam, Korea).

Techniques: Control

Journal: Annals of Dermatology

Article Title: Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis

doi: 10.5021/ad.2019.31.2.186

Figure Lengend Snippet: Effects of adiponectin on expression of lipid synthesis related gene in atopic dermatitis reconstructed epidermal model. By using the quantitative real-time reverse transcriptase polymerase chain reaction analysis, major rate-limiting epidermal lipid biosynthesis enzymes (A) fatty acid synthase, (B) HMG CoA reductase, and (C) serine palimitoyl transferase, and the transcription factors encoding for lipid synthesis enzymes (D) sterol regulatory binding protein 1a (SREBP1a), (E) SREBP1c, and (F) SREBP2 were assessed. * p <0.05, ** p <0.01.

Article Snippet: Recombinant human full-length adiponectin protein was obtained from Biobud (Seongnam, Korea).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Binding Assay

Journal: Annals of Dermatology

Article Title: Adiponectin Attenuates the Inflammation in Atopic Dermatitis-Like Reconstructed Human Epidermis

doi: 10.5021/ad.2019.31.2.186

Figure Lengend Snippet: Effects of adiponectin on epidermal differentiation in atopic dermatitis (AD) reconstructed epidermal model. The mRNA expression of the major epidermal differentiation markers (A) filaggrin (FLG), (B) loricrin, and (C) involucrin was quantified using real time reverse transcriptase polymerase chain reaction. (D) Representative immunohistochemical staining of untreated normal epidermal equivalent (normal-HEEs), and AD mimicking epidermis equivalent (AD-HEEs), and adiponectin treated AD mimicking equivalent (AD-HEEs+adiponectin) with human filaggrin antibody. Tx.: treatment. Bar=50 µm. * p <0.05, ** p <0.01.

Article Snippet: Recombinant human full-length adiponectin protein was obtained from Biobud (Seongnam, Korea).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Immunohistochemical staining, Staining