Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 1: Effect of oral intake of Plaquenil on GDF15 and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Clinical Proteomics, Transformation Assay

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 2: Distribution and characterization of GDF15 expressing epithelial cells in the proximal gut of normal-weight individuals and patients with obesity. A) Relative mRNA expression (efficiencyDDCt method) of GDF15 in resection specimens from the fundus (nNW ¼ 7, nOB ¼ 10), corpus (nNW ¼ 5, nOB ¼ 8), antrum (nNW ¼ 5, nOB ¼ 8), and jejunum (nNW ¼ 5, nOB ¼ 7) of normal-weight individuals and patients with obesity. (Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 in jejunal sections (10 mm) of a normal-weight individual and a patient with obesity. GDF15þ cells were stained with Cy3 (green) and nuclei were stained with DAPI (blue). Scale bars: 25 mm. C) Average number and intensity of GDF15þ cells in jejunal tissue sections from both normal-weight individuals and patients with obesity (nNW ¼ 5, nOB ¼ 5; two-tailed unpaired Student’s t-test). DeH) Representative double-immunofluorescence staining for GDF15 (green [Cy3 or Alexa594]) with markers for (D) goblet (MUC2: red [Cy5]), (E) Paneth (a-defensin 6: red [Cy5]) or (FeG) enteroendocrine (CHGA: red [Alexa488]) and ghrelin (red [Alexa488]) cells in jejunal sections (10 mm) from normal-weight individuals. Arrows indicate co-localization. Normal rabbit serum was used as negative control. Nuclei were labeled with DAPI (blue). Scale bars: 25 mm. NW: normal-weight, OB: obese. Data of figure A, C represent mean SEM and single values are plotted. *P < 0.05: versus jejunum in normal-weight individuals, $$$P < 0.001: versus jejunum in patients with obesity, #P < 0.05, ###P < 0.001: versus normal-weight individuals.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Staining, Two Tailed Test, Negative Control, Labeling

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 3: Effects of bitter generalists and intermediates on GDF15 or GLP-1 expression in jejunal crypts from patients with obesity. A) Effect of 4-hour stimulation with 100 mM HCQS on relative GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). B) Effect of 4-hour stimulation with 100 mM HCQS on relative GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). C) Effect of bitter generalists on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 6e9; Proc Mixed Model with Sidák correction for multiple comparisons). D) Effect of bitter intermediates on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e6; Proc Mixed Model with Sidák correction for multiple comparisons). E) Relative GDF15 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin. Azithromycin was removed after 4 h for the 24-hour timepoint (n ¼ 3e6; one-tailed paired Student’s t-test). F) Relative GLP-1 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin for 6 h (n ¼ 3; two-tailed paired Student’s t-test). All data are present as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus the vehicle group.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Two Tailed Test, One-tailed Test

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 4: Effects of bitter specialists on GDF15 mRNA or protein expression in primary jejunal crypts from patients with obesity. A) Effect of specialists on relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e7; Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 (green [Cy3]) in primary crypts treated with 1 mM gallic acid or vehicle after 24 h with 4 h stimulation. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. C) Average intensity of GDF15þ cells in primary crypts treated with 1 mM gallic acid or vehicle. Gallic acid was removed after 4 h of stimulation and staining was performed 20 h later (n ¼ 4; two-tailed paired Student’s t-test). D) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen for 6 h (n ¼ 3; one-tailed paired Student’s t-test). E) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen. Acetaminophen was removed after 4 h for the 24-hour timepoint (n ¼ 3; one-tailed paired Student’s t-test). F) Venn diagram of bitter compounds that affected GDF15 and/or GLP-1 mRNA expression after stimulation or primary crypts for 4 h. Asterisk (*) indicates an inhibitory effect on mRNA expression; the use of hashtags (#) signifies potential variations in the effects on GDF15 or GLP-1 mRNA, depending on the genotype of the patients. Data of figure A, C, D, E are present as mean SEM and single values are plotted. **P < 0.01, ***P < 0.001: versus the vehicle group.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Staining, Labeling, Two Tailed Test, One-tailed Test

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 5: Role of TAS2Rs and the motilin receptor in the effect of bitter agonists on GDF15 or GLP-1 expression in primary crypts from patients with obesity. A) The TAS2R antagonist, GIV3727 (110 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). B) GIV3727 (110 mM) did not block the effect of gallic acid (1 mM) on GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). C) GIV3727 (110 mM) did not block the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). D) The motilin receptor antagonist MA-2029 blocked the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). E) C12-O-AHL (0.15 mM) increased relative GDF15 mRNA expression in primary crypts from obese patients with TAS2R4(GG/CG) (n ¼ 9) but not TAS2R4(CC) (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). F) C12-O-AHL (0.15 mM) decreased GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R4(GG/CG) (n ¼ 9) but not the TAS2R4(CC) genotype (n ¼ 4). (Proc Mixed Model with Sidák correction for multiple comparisons). G) Aloin at 30 mM (white dots) and 100 mM (black dots) decreased relative GDF15 mRNA expression in primary crypts from obese patients with the TAS2R43(þ)GG/CG (n ¼ 9) but not the TAS2R43(þ)CC (n ¼ 6) or TAS2R43() (n ¼ 6) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). H) Aloin at 30 mM (white dots) and 100 mM (black dots) did not affect the relative GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R43(þ) GG/GC (n ¼ 9), TAS2R43(þ) CC (n ¼ 6), or TAS2R43() (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). IeK) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in jejunal sections (10 mm thickness) from normal-weight populations. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. MeO) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in primary jejunal crypt from patients with obesity. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. (L and P) Normal rabbit serum as a negative control. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. Data of figure AeH are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01, ###P < 0.001: bitter treatment antagonist, $P < 0.05: versus TAS2R4(CC) population, &P < 0.05: versus TAS2R43() population.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing, Blocking Assay, Labeling, Negative Control

Journal: Molecular metabolism

Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

doi: 10.1016/j.molmet.2024.102002

Figure Lengend Snippet: Figure 6: Role of the unfolded protein response pathway in the effect of bitter agonists on GDF15 and GLP1 mRNA expression in primary crypts from patients with obesity. A) Positive correlation between Log2 fold change of GDF15 and DDIT3 mRNA expression induced by bitter agonists (bitter: DB, AZI, GA, PHE, EM, ACE, BERB; n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). B) Positive correlation between Log2 fold change of GDF15 and ATF4 mRNA expression induced by bitter agonists from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). C) Positive correlation between Log2 fold change of DDIT3 and ATF4 mRNA expression induced by bitter agonist from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). D) ISRIB (5 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). E) ISRIB (5 mM) did not affect the effects of gallic acid (1 mM) on GLP-1 mRNA expression (n ¼ 2; Proc Mixed Model with Sidák correction for multiple comparisons). Data are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01: bitter treatment antagonist.

Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

Techniques: Expressing

Journal: Open Access Macedonian Journal of Medical Sciences

Article Title: Impact of the Neuregulin rs35753505 C/T Polymorphisms on Neuregulin 1 Levels in Preterm Infants

doi: 10.3889/oamjms.2019.554

Figure Lengend Snippet: The differences in serum NRG1 levels between genotypes and alleles of NRG rs35753505 C/T polymorphisms

Article Snippet: The association between polymorphisms and NRG1 levels was analysed using the chi-square test.

Techniques:

Journal: Open Access Macedonian Journal of Medical Sciences

Article Title: Impact of the Neuregulin rs35753505 C/T Polymorphisms on Neuregulin 1 Levels in Preterm Infants

doi: 10.3889/oamjms.2019.554

Figure Lengend Snippet: The association between NRG rs35753505 C/T polymorphism with NRG1 levels

Article Snippet: The association between polymorphisms and NRG1 levels was analysed using the chi-square test.

Techniques:

Journal: bioRxiv

Article Title: Early neuroadaptations to an obesogenic diet identify the schizophrenia-related ErbB4 receptor in obesity-induced hippocampal abnormalities

doi: 10.1101/2021.06.30.450398

Figure Lengend Snippet: (A) Study 1 timeline and study groups. (B) Study 1 timeline of behavioral procedures, MRI, and histology. (C) Study 2 timeline of molecular analyses. Abbreviations: CD , control diet; EPM , elevated plus maze; FPS , fear-potentiated startle; MRI , magnetic resonance imaging; NRG1 , neuregulin-1; OFT , open field test; PND , postnatal day; VEH , vehicle; WD , Western diet.

Article Snippet: NRG1 levels were determined using the SpectraMax i3X plate reader (Molecular Devices, Sunnyvale, CA USA) as the mean absorbance at 450nm with correction at 540 nm using a standard curve of samples ranging from 0 to 4,000 pg/mL.

Techniques: Magnetic Resonance Imaging, Western Blot

Journal: bioRxiv

Article Title: Early neuroadaptations to an obesogenic diet identify the schizophrenia-related ErbB4 receptor in obesity-induced hippocampal abnormalities

doi: 10.1101/2021.06.30.450398

Figure Lengend Snippet: (A-B) Total hippocampal volume (both left and right hippocampi combined) were affected by the treatment [ F (1, 19) = 9.48, p = .006]. Subchronic NRG1 administration during adolescence resulted in significant total hippocampal volume reduction (CDV vs. CDN, p = .003). (C-D) Hippocampal subfield segmentation showed a significant interaction [ F (1, 19) = 6.44, p = .020] effects on CA1 region. WDV rats had a significantly smaller CA1 volume than CDV rats ( p = .015). Sample size = 6 rats / group.

Article Snippet: NRG1 levels were determined using the SpectraMax i3X plate reader (Molecular Devices, Sunnyvale, CA USA) as the mean absorbance at 450nm with correction at 540 nm using a standard curve of samples ranging from 0 to 4,000 pg/mL.

Techniques:

Journal: bioRxiv

Article Title: Early neuroadaptations to an obesogenic diet identify the schizophrenia-related ErbB4 receptor in obesity-induced hippocampal abnormalities

doi: 10.1101/2021.06.30.450398

Figure Lengend Snippet: Full-focused composite 40x images were acquired from twelve (12) rats. Hippocampal dorsal and ventral CA1 areas were scanned to obtain at least three (3) images per region of interest per hemisphere covering a field of view area of 1.2 mm. Each acquired image was saved as a TIFF file and included at least six (6) Iba-1 + cells. (A) Representative microglia morphologies from hippocampal dorsal and ventral CA1 regions in each study group. (B) Evaluation of microglial density as a measure of cell size based on area demonstrated that the WD significantly decreased microglial density in the right CA1 subfield (diet: [ F (1,8) = 6.82, p < .031]). (C) Lacunarity was significantly decreased in the right CA1 subfield of rats that consumed an obesogenic WD ( diet: [ F (1,8) = 12.09, p < .008]). The span ratio was significantly increased in the right CA1 region of rats receiving the exogenous NRG1 ( treatment: [ F (1,8) = 15.43, p < .004]). NRG1 administration reduced circularity in the right CA1 region ( treatment: [ F (1,8) = 9.67, p < .014]). Sample size = 3 rats / group.

Article Snippet: NRG1 levels were determined using the SpectraMax i3X plate reader (Molecular Devices, Sunnyvale, CA USA) as the mean absorbance at 450nm with correction at 540 nm using a standard curve of samples ranging from 0 to 4,000 pg/mL.

Techniques:

Journal: bioRxiv

Article Title: Early neuroadaptations to an obesogenic diet identify the schizophrenia-related ErbB4 receptor in obesity-induced hippocampal abnormalities

doi: 10.1101/2021.06.30.450398

Figure Lengend Snippet: Bead-based flow cytometry immunoassay was used to evaluate cytokine levels. (A-D) The WD led to a significant increase in cytokines in the hippocampus: IL1-α ( p = .010), TNF-α ( p = .006), IL-10 ( p = .027), and IL-6 ( p = .019). (E-H) NRG1 administration significantly reduced IL-33 ( p = .041), GM-CSF ( p = .021), CCL-2 ( p = .037), and IFN-γ ( p = .031). includes the detailed statistics of the cytokines listed in . Sample size = 5-6 rats / group.

Article Snippet: NRG1 levels were determined using the SpectraMax i3X plate reader (Molecular Devices, Sunnyvale, CA USA) as the mean absorbance at 450nm with correction at 540 nm using a standard curve of samples ranging from 0 to 4,000 pg/mL.

Techniques: Flow Cytometry

Journal: bioRxiv

Article Title: Early neuroadaptations to an obesogenic diet identify the schizophrenia-related ErbB4 receptor in obesity-induced hippocampal abnormalities

doi: 10.1101/2021.06.30.450398

Figure Lengend Snippet: (A) Obesogenic WD decreased total ErbB4 protein levels in the hippocampus (diet [ F (1, 16) = 7.34, p = .016]). WDN rats exhibited significantly lower ErbB4 protein levels when compared to CDV rats ( p = .009). (B) Phosphorylation of hippocampal ErbB4 protein levels was affected by the treatment [ F (1, 18) = 5.82, p = .027] and diet [ F (1,18) = 7.36, p = .014]. ErbB4 hyperphosphorylation was particularly evident in WDN rats relative to the CDV group ( p = .012). (C) TACE/ADAM17 protein levels in the hippocampus were affected by the treatment [ F (1, 17) = 8.93, p = .008] and diet [ F (1, 17) = 11.03, p = .004]. WDN rats showed increased TACE/ADAM17 protein levels when compared to CDV rats ( p = .001). (D) NRG1 administration showed a significant decrease for PSD-95 protein levels in the hippocampus (treatment [ F (1, 18) = 4.47, p = .048]). Sample size = 5-6 rats / group.

Article Snippet: NRG1 levels were determined using the SpectraMax i3X plate reader (Molecular Devices, Sunnyvale, CA USA) as the mean absorbance at 450nm with correction at 540 nm using a standard curve of samples ranging from 0 to 4,000 pg/mL.

Techniques: