Journal: Molecular medicine reports

Article Title: Function of ghrelin and ghrelin receptors in the network regulation of gastric motility.

doi: 10.3892/mmr.2014.2571

Figure Lengend Snippet: Figure 3. GHS-R1s and α-actin immunofluorescent staining in cultured smooth muscle cells. (A) GHS-R1s staining (red fluorescence, arrows); (B) α-actin staining (green fluorescence, arrows); (C) GHS-R1s/α-actin staining (merged image fluorescence, arrows). Cell nuclei, DAPI staining. Scale bar, 15 µm (magnification, x40). GHS-Rs, ghrelin receptors.

Article Snippet: Rat ghrelin, [d-Lys3]-GHRP-6 (an antagonist of GHS-R), atropine (an antagonist of M-type cholinergic receptor), carbamoylcholine chloride and nimodipine were obtained from Tocris Cookson (Bristol, UK), tetrodotoxin (TTX) was obtained from Sigma (St. Louis, MO, USA), goat anti-rat GHS-R1 (D-16) antibody, mouse anti-rat NF-H (H-5) antibody, mouse anti-rat a-actin antibody (1A4) antibody and rabbit anti-rat c-Kit antibody (C-19) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies as well as tetramethylrhodamine (TRITC)-conjugated rabbit anti-goat secondary antibody were obtained from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).

Techniques: Staining, Cell Culture, Fluorescence

Journal: Molecular medicine reports

Article Title: Function of ghrelin and ghrelin receptors in the network regulation of gastric motility.

doi: 10.3892/mmr.2014.2571

Figure Lengend Snippet: Figure 1. GHS-R1s and NF-H immunofluorescent staining in gastric antrum nerve cells. (A) GHS-R1s staining (red fluorescence; big arrows indicate cellular bodies and small arrows indicate neural fibers); (B) NF-H staining (green fluorescence, arrows); (C) GHS-R1s/NF-H staining (merged image fluorescence, big arrows indicate cellular bodies and small arrows indicate neural fibers). Cell nuclei, DAPI staining. Scale bar, 15 µm (magnification, x40). GHS-Rs, ghrelin receptors; NF-H, neurofilament heavy polypeptide.

Article Snippet: Rat ghrelin, [d-Lys3]-GHRP-6 (an antagonist of GHS-R), atropine (an antagonist of M-type cholinergic receptor), carbamoylcholine chloride and nimodipine were obtained from Tocris Cookson (Bristol, UK), tetrodotoxin (TTX) was obtained from Sigma (St. Louis, MO, USA), goat anti-rat GHS-R1 (D-16) antibody, mouse anti-rat NF-H (H-5) antibody, mouse anti-rat a-actin antibody (1A4) antibody and rabbit anti-rat c-Kit antibody (C-19) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies as well as tetramethylrhodamine (TRITC)-conjugated rabbit anti-goat secondary antibody were obtained from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).

Techniques: Staining, Fluorescence

Journal: Molecular medicine reports

Article Title: Function of ghrelin and ghrelin receptors in the network regulation of gastric motility.

doi: 10.3892/mmr.2014.2571

Figure Lengend Snippet: Figure 2. GHS-R1s and c-Kit immunofluorescent staining in gastric antrum Cajal cells. (A) GHS-R1s staining (red fluorescence, arrows); (B) c-Kit staining (green fluorescence, big/small arrows); (C) GHS-R1s/c-Kit staining (merged image fluorescence, big arrows). Scale bar, 20 µm (magnification, x40). GHS-Rs, ghrelin receptors.

Article Snippet: Rat ghrelin, [d-Lys3]-GHRP-6 (an antagonist of GHS-R), atropine (an antagonist of M-type cholinergic receptor), carbamoylcholine chloride and nimodipine were obtained from Tocris Cookson (Bristol, UK), tetrodotoxin (TTX) was obtained from Sigma (St. Louis, MO, USA), goat anti-rat GHS-R1 (D-16) antibody, mouse anti-rat NF-H (H-5) antibody, mouse anti-rat a-actin antibody (1A4) antibody and rabbit anti-rat c-Kit antibody (C-19) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies as well as tetramethylrhodamine (TRITC)-conjugated rabbit anti-goat secondary antibody were obtained from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).

Techniques: Staining, Fluorescence

Journal: Open Life Sciences

Article Title: Physical activity enhances fecal lactobacilli in rats chronically drinking sweetened cola beverage

doi: 10.1515/biol-2022-0070

Figure Lengend Snippet: Effect of SPA on insulin, adiponectin, and ghrelin levels in plasma of controls and cola SSB drinking rats. CON = control group, SSB = group drinking sugar sweetened beverage, SSB + SPA = group drinking sugar sweetened beverage concomitant to spontaneous physical activity, SPA = group performing spontaneous physical activity. Average ± SEM. n = 8 per group; * p < 0.05 vs SSB, # p < 0.05 vs CON.

Article Snippet: To measure and analyze adiponectin (as an adipose tissue-related indicator of metabolic changes), insulin (as a pancreas-related indicator of metabolic changes), and ghrelin (as a gastrointestinal tract-related indicator of appetite), commercially available kits for ELISA were used (Rat Ghrelin ELISA Kit, Elabscience, USA; Rat Adiponectin ELISA Kit, Abcam, UK; and Rat Insulin ELISA Kit, Mercodia, Sweden, respectively).

Techniques: Clinical Proteomics, Control, Activity Assay

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: Unacylated ghrelin (UAG; 100pM) inhibits the growth of ( a ) MCF7 and ( b ) MDA-MB-468 cells grown in 3D in matrigel or collagen, but not in 2D. ( c ) UAG had no effect on MDA-MB-231 cell growth in 2D or 3D. Dose-dependent effects of UAG and doxorubicin on ( d ) MCF7, ( e ) MDA-MB-468 and ( f ) MDA-MB-231 cell number in 3D culture. ( g ) Acyl ghrelin (AG) and UAG (100 pM) inhibit breast cancer cell growth in serum-stimulated conditions. ( h ) Binding of Cy3-labeled UAG to MCF7, MDA-MB-468 and MDA-MB-231 cells over time (representative images). Scale bar represents 20 μm. Data represent mean ± SEM with three replicates/group. Experiments repeated at least twice. UAG: unacylated ghrelin; AG: acylated ghrelin; VC: vehicle control; FBS: fetal bovine serum. Figure 1—figure supplement 1—source data 1. Unacylated ghrelin inhibits the growth of breast cancer cells in 3D.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Binding Assay, Labeling, Control

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: ( a, c ) Unacylated ghrelin (UAG; 100 pM) inhibits the growth of a panel of breast cancer cell lines under serum-stimulated conditions (six replicates/group) or ( b ) ER+ breast cancer cell lines in the presence of estradiol (10 nM; six replicates/group). ( c ) UAG (100 pM) suppresses cell growth of basal-like and mesenchymal-like TNBC breast cancer cell lines that are WT for BRAF and KRAS (6–9 replicates/group). Effects of UAG are abrogated in ( d ) BRAF-transfected MCF7 cells, and ( e ) BRAF - and ( f ) KRAS -mutated colon cancer cells (6–12 replicates/group). Loss of mutated alleles of BRAF or KRAS sensitizes cells to the effect of UAG. Data represent mean ± SEM. Experiments were repeated at least twice. UAG: unacylated ghrelin; VC: vehicle control; FBS: fetal bovine serum; E2: estradiol. Figure 1—source data 1. Unacylated ghrelin inhibits the 3D growth of breast cancer cells.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Transfection, Control

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: Characteristics of breast cancer cell lines and patient-derived breast cancer cells, and responsiveness to unacylated ghrelin.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques:

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: ( a ) Unacylated ghrelin (UAG; 10–1000 pM) inhibits the forskolin-stimulated production of cAMP in MCF7 cells (3–4 replicates/group). ( b ) UAG (100 pM) stimulates activation of Gαi (three replicates/group). ( c ) UAG (100 pM) suppresses the growth of CRISPR GNAI1 and GNAI3 KO cells, but not GNAI2 KO MCF7 cells, suggesting Gαi2-coupled GPCR-mediated effects. Suppression of ( d ) estradiol- or ( e ) serum-stimulated breast cancer cell growth with UAG (100 pM) is prevented in the presence of Gαi inhibitor, pertussis toxin (20 ng/ml, 200 ng/ml; three replicates/group). ( f ) PKA inhibitor (KT5720), adenylyl cyclase inhibitor (SQ22536) or cAMP antagonist (cAMPS-RP) suppress the serum-stimulated growth of MCF7 cells (three replicates/group). ( g ) UAG (100 pM) inhibits the forskolin-stimulated growth of MCF7 and MDA-MB-468 cells, but not MDA-MB-231 (three replicates/group). ( h ) U0126 (MEK inhibitor) inhibits the forskolin-stimulated growth of MCF7, MDA-MB-468 and MDA-MB-231 cells (three replicates/group). ( i ) A model summarizing the putative mechanism of action of UAG in breast cancer cells and compounds used to dissect mechanism of action. Data represent mean ± SEM. Experiments were repeated at least twice. UAG: unacylated ghrelin; VC: vehicle control; FBS: fetal bovine serum; E2: estradiol; PT: pertussis toxin; M: melatonin; FSK: forskolin. Figure 2—source data 1. Unacylated ghrelin suppresses breast cancer cell growth via Gαi-dependent inhibition of cAMP formation. Figure 2—source data 2. Unacylated ghrelin suppresses breast cancer cell growth via Gαi-dependent inhibition of cAMP formation.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Activation Assay, CRISPR, Control, Inhibition

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: ( a ) Unacylated ghrelin (UAG) has no effect on the release of intracellular Ca2+ in MCF7 cells. ( b ) Loss of GNAI2 (blue outline) using CRISPR and three validated gRNAs is associated with loss of response to UAG (100 pM), while KO of GNAI1 or GNAI3 had no effect (green and red outline, respectively). ( c ) Suppression of estradiol-stimulated ZR75 cell growth by UAG (100 pM) is prevented by Gαi inhibitor, pertussis toxin (20 ng/ml, 200 ng/ml). ( d ) PKA (KT5720) or adenylyl cyclase (SQ22536) inhibitors, or cAMP antagonist (cAMPS-RP) suppress the growth of MDA-MB-468 cells. ( e ) UAG (100 pM) inhibits the forskolin-stimulated growth of ZR75 cells. Data represent mean ± SEM with three replicates/group. Experiments were repeated at least twice. UAG: unacylated ghrelin; FBS: fetal bovine serum; E2: estradiol; FSK: forskolin. Figure 2—figure supplement 1—source data 1. Unacylated ghrelin suppresses breast cancer cell growth via Gαi-dependent mechanisms.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: CRISPR

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: ( a ) Unacylated ghrelin (UAG; 100 pM) inhibits ERK activity (EYFP FRET) in EKAR-transfected MCF7 cells (10 replicates/group). Data were normalized to vector ECFP signal. Scale bar represent 50 μm. Western blotting demonstrates that UAG causes a decrease in the ( b ) phosphorylation of ERK1/2 and downstream MAPK target p90RSK and ( c ) expression of cMYC in MCF7 cells. UAG also causes a decrease in ( d ) the phosphorylation of Akt and its downstream target, p70S6K, as well as ( e ) FoxO3a nuclear localization FoxO3a-RFP-transfected cells, an effect that is attenuated in cells treated with PI3K inhibitor LY294002 (five replicates/group). Data represent mean ± SEM. Experiment were repeated at least twice. UAG: unacylated ghrelin; FBS: fetal bovine serum. Figure 3—source data 1. Unacylated ghrelin suppresses EKAR and FoxO3 nuclear localization. Figure 3—source data 2. Unacylated ghrelin inhibits MAPK and Akt signaling.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Activity Assay, Transfection, Plasmid Preparation, Western Blot, Phospho-proteomics, Expressing

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: Unacylated ghrelin (UAG) significantly inhibits the proliferation of ( a ) MCF7, ( b ) ZR75 and ( c ) MDA-MB-468 in the presence of estradiol (10 nM) or serum (3–6 replicates/group). Representative images showing EdU incorporation (green). Hoechst nuclear stain; blue. Scale bar represent 100 μm. Effects are mediated via induction of G1-phase cell cycle arrest (RFP+) and a reduction in the number of cells in S/G2/M-phase (GFP+) and G1/S transition (YFP+) in ( d ) MCF7 and ( e ) MDA-MB-468 cells (four replicates/group). (Hoechst nuclear stain; blue). Scale bar represents 100 μm. ( f ) UAG stimulates cell death in MCF7 and MDA-MB-468 cells, but not MDA-MB-231 cells (three replicates/group). Western blot results demonstrating that UAG inhibits CDK4/cyclin D3, pRB (Ser 795) and BCL2, and stimulates BAX in ( g ) MCF7 and ( h ) MDA-MB-468 cells. Data represent mean ± SEM. Experiments were repeated at least twice. UAG: unacylated ghrelin; VC: vehicle control; FBS: fetal bovine serum; E2: estradiol. Figure 4—source data 1. Unacylated ghrelin causes cell cycle arrest and apoptosis. Figure 4—source data 2. Unacylated ghrelin causes cell cycle arrest and apoptosis.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Staining, Western Blot, Control

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: ( a ) Unacylated ghrelin (UAG) significantly inhibits the proliferation of tamoxifen-resistant LCC2 cells in the presence of estradiol (10 nM). Representative images showing EdU incorporation (green). Hoechst nuclear stain (blue). Scale bar represents 100 μm. ( b ) No effect of UAG on cell cycle was observed in MDA-MB-231 cell using FUCCI cell cycle system. Flow cytometry analysis demonstrating that UAG induces G1-phase cell cycle arrest and apoptosis in ( c, d ) MCF7, ( e, f ) MDA-MB-468 and ( g, h ) ZR75 cells. Data represent mean ± SEM with three replicates/group. Experiments were repeated at least twice. UAG: unacylated ghrelin; FBS: fetal bovine serum; E2: estradiol. Figure 4—figure supplement 1—source data 1. Unacylated ghrelin causes cell cycle arrest and apoptosis.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Staining, Flow Cytometry

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: Tumor volume in response to treatment with 50 μg/kg (blue), 100 μg/kg (red) or 200 μg/kg (purple) UAG in mice xenografted with ( a ) MCF7 (six replicates/group), ( b ) ZR75 (five replicates/group), or allografted with ( c ) J110 (five replicates/group) cells. Representative tumor (below) with scale bar representing 10 mm. UAG significantly increases the number of cells with apoptotic nuclei in ( d ) MCF7, ( e ) ZR75 and ( f ) J110 xenografts. ( g ) UAG (100 pM) significantly inhibits the growth of patient-derived ER+ breast cancer cells and 4013-TG3 TNBC cells, but not 3204-TG6 TNBC cells. ( h ) Heatmap representing baseline differential expression of MAPK-target genes in responsive vs. non-responsive patient-derived cells. Data represent mean ± SEM. UAG: unacylated ghrelin; VC: vehicle control; FBS: fetal bovine serum. Figure 5—source data 1. Unacylated ghrelin and cyclic analog AZP-531 inhibit tumor growth in xenograft models and patient-derived tumor cells.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Derivative Assay, Quantitative Proteomics, Control

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: Representative images of tumors in response to treatment with 50 μg/kg or 100 μg/kg unacylated ghrelin (UAG) in nude mice xenografted with ( a ) MCF7 (six replicates/group) and ( b ) ZR75 (five replicates/group) cells. Effect of UAG on tumor volume of J110 (five replicates/group) ( c ) allografts in FVB mice or ( d ) xenografts in nude mice compared to vehicle control. ( e ) UAG (100 pM) significantly inhibits the growth of patient-derived ER+ breast cancer, 2147-TG5 TNBC, but has no effect on the growth of 3887-TG7 TNBC. Data represent mean ± SEM. Representative images of tumors in response to treatment with 200 μg/kg AZP-531 in ( f ) NSG mice xenografted with MDA-MB-468 cells (8–9 replicates/group) or ( g ) FVB mice allografted with J110 cells (five replicates/group). Scale bars represent 10 mm. UAG: unacylated ghrelin; VC: vehicle control. Figure 5—figure supplement 1—source data 1. Unacylated ghrelin and cyclic analog AZP-531 inhibit tumor growth in xenograft models andpatient-derived tumor cells.

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Control, Derivative Assay

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet: AZP-531 causes the dose-dependent inhibition of ( a ) MCF7 and ( b ) MDA-MB-468 and ( c ) patient-derived TNBC breast cancer cell growth in 3D, compared with chemotherapeutic agent doxorubicin (three replicates/group). ( d ) Unacylated ghrelin (UAG; 10–1000 pM) and AZP-531 (AZP; 10–1000 pM) inhibits the forskolin-stimulated production of cAMP in MCF7 cells (3–6 replicates/group). Data represent mean ± SEM. Experiments were repeated at least twice. Tumor volume in response to treatment with 200 μg/kg AZP-531 (purple) in mice xenografted with ( e ) MDA-MB-468 (8–9 replicates/group) or allografted with ( f ) J110 (five replicates/group) cells. Representative images (below) with scale bars representing 10 mm. Figure 6—source data 1. Unacylated ghrelin analog, AZP-531, inhibits breast cancer cell growth in vitro , ex vivo and in vivo .

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Inhibition, Derivative Assay, In Vitro, Ex Vivo, In Vivo

Journal: eLife

Article Title: Three-dimensional growth of breast cancer cells potentiates the anti-tumor effects of unacylated ghrelin and AZP-531

doi: 10.7554/eLife.56913

Figure Lengend Snippet:

Article Snippet: Peptide, recombinant protein , [Des-octanoyl]-Ghrelin (rat) , Tocris Bioscience , Cat# 2951 , .

Techniques: Recombinant, Plasmid Preparation, Sequencing, Imaging, Activation Assay, Software, Microscopy, Membrane

Journal: Endocrinology

Article Title: Molecular forms of hypothalamic ghrelin and its regulation by fasting and 2-deoxy-d-glucose administration.

doi: 10.1210/en.2005-0174

Figure Lengend Snippet: FIG. 3. A, Body weights (left panel) and blood glucose concentrations (right panel) of rats after fasting. B, Ghrelin mRNA levels in the hypothalamus and stomach. C, NPY, AgRP, and MCH mRNA levels in the hypothalamus of rats fed ad libitum (control) or animals that fasted for 24 or 48 h. Note that the values of the longitudinal axes are different in each graph. Asterisks indicate the differences between each bar (P 0.05).

Article Snippet: Real-time PCR of rat ghrelin, NPY, AgRP, and melaninconcentrating hormone (MCH) Total RNA was extracted from frozen rat stomach and hypothalamus samples using TRIzol (Invitrogen, Tokyo, Japan).

Techniques: Control

Journal: Endocrinology

Article Title: Molecular forms of hypothalamic ghrelin and its regulation by fasting and 2-deoxy-d-glucose administration.

doi: 10.1210/en.2005-0174

Figure Lengend Snippet: FIG. 5. A, Food intake in the 2 h after 2-DG injection. B, Ghrelin mRNA levels in the hypothalamus and stomach. C, NPY, AgRP, and MCH mRNA levels in the hypothalami of rats treated with 2-DG for 2 h. Asterisks indicate the differences between each bar (P 0.05).

Article Snippet: Real-time PCR of rat ghrelin, NPY, AgRP, and melaninconcentrating hormone (MCH) Total RNA was extracted from frozen rat stomach and hypothalamus samples using TRIzol (Invitrogen, Tokyo, Japan).

Techniques: Injection

Journal: International journal of molecular sciences

Article Title: Maternal Exposure to Low-Dose BDE-47 Induced Weight Gain and Impaired Insulin Sensitivity in the Offspring.

doi: 10.3390/ijms25168620

Figure Lengend Snippet: Figure 2. Maternal BDE-47 exposure and expression of key genes in adipose tissue and liver and serum adipokines in the female offspring. (A) The mRNA expression levels of selected target genes investigated in adipose tissue and (B) liver of 12-week-old female offspring. (C) Levels of leptin, adiponectin, resistin, and ghrelin measured in serum of female offspring. Data are expressed as mean ± SEM, n ≥5. Significant differences were derived by Mann–Whitney test with * p < 0.05.

Article Snippet: Adiponectin, leptin, resistin, and acetylated ghrelin serum concentrations were determined by ELISA using mouse standards according to the manufacturer’s guidelines (mouse adiponectin, leptin, resistin ELISA; R&D Systems, Minneapolis, MN, USA), (mouse/rat acylated ghrelin ELISA; BioVendor, Karasek, Czech Republic).

Techniques: Expressing, Derivative Assay, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Maternal Intact Protein- and Amino Acid-Based Diets on Development of Food Intake Regulatory Systems and Body Weight in Dams and Male Offspring of Wistar Rats

doi: 10.3390/ijms20071690

Figure Lengend Snippet: Effect of maternal diet on fasting plasma ghrelin, PYY, glucose, and insulin in dams at week 9 post-partum and in pups at birth, weaning, and at week 18 post-weaning. ( n = 5–6/group). Data are means ± SEM; AAD: amino acid-based diet; IPD: intact protein diet; PYY: Peptide YY.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) were used to measure plasma concentrations of PYY (catalog no. 48-PYYRT-E01.1, Alpco Diagnostics, Salem, NH, USA), ghrelin (catalog no. 32-5118, Alpco Diagnostics), and insulin (catalog no. 80-INSRT-E01, Alpco Diagnostics).

Techniques: Clinical Proteomics

Journal: NPJ biofilms and microbiomes

Article Title: Yijung-tang improves thermogenesis and reduces inflammation associated with gut microbiota in hypothyroid rats.

doi: 10.1038/s41522-023-00396-2

Figure Lengend Snippet: Fig. 1 Effects of Yijung-tang (YJT) and L-thyroxine (T4) in metabolic phenotypes, serum metabolites, and histology in hypothyroid rats. a Schematic overview of the experimental design; b body mass; c food intake; d average core body temperature (Tb) during the experiment (n = 5 per group). e Glucose metabolism was assessed via an oral glucose tolerance test. f–hThe levels of thyroid stimulating hormone (TSH), thyroxine (T4), and tri-iodothyronine (T3). i–l Serum ghrelin, glucagon-like peptide-1 (GLP-1), tumor necrosis factor-α (TNF-α), and lipopolysaccharides (LPS). m Histology assay for the ileum by hematoxylin and eosin for four experimental groups. Data are presented as means ± SEM (n = 7–8 per group). One-way ANOVA or ANCOVA (food intake) followed by post hoc LSD test. *P < 0.05, versus control, #P < 0.05 versus PTU. Different letters above the columns indicate significant differences. Con, control group that received only saline; PTU, the rats that received 10 mg/kg/body propylthiouracil (PTU); YJT + PTU, the rats that received 2.1 g/kg YJT and 10 mg/kg/body PTU; T4 + PTU, the rats that were treated with 0.5 mg/kg L-thyroxine and 10 mg/kg PTU.

Article Snippet: Published in partnership with Nanyang Technological University npj Biofilms and Microbiomes (2023) 32 Serum biochemical parameter assays We used enzyme-linked immunosorbent assay kits (ELISA) to estimate the levels in the serum, including TSH (CSB-E05115r), free Tri-iodothyronine (free-T3, CSB-E05076r), thyroxine (T4, CSBE05082r), GLP-1 (CSB-E08117r), TNF-α (CSB-E11987r), ghrelin (CSB-E09816r), and LPS (CSB-E14247r) following the manufacturer’s recommendations (Cusabio, Wuhan, China).

Techniques: Control, Saline

Journal: NPJ biofilms and microbiomes

Article Title: Yijung-tang improves thermogenesis and reduces inflammation associated with gut microbiota in hypothyroid rats.

doi: 10.1038/s41522-023-00396-2

Figure Lengend Snippet: Fig. 4 Cecal microbiota transfer (CMT) modulates the metabolic phenotypes, serum metabolites, and ileum histology in recipients. a Schematic overview of the experimental design. b Average core body temperature (Tb) during the experiment process (n = 4 per group). c Blood glucose levels. d–h The serum thyroxine (T4), tri-iodothyronine (T3), ghrelin, glucagon-like peptide-1 (GLP-1), and lipopolysaccharides (LPS) by ELISA. i Histology assay for the ileum by hematoxylin and eosin staining for four experimental groups. The scale bar is 500 µm. The data are presented as means ± SEM. One-way ANOVA followed by post hoc LSD test. *P < 0.05 vs vehicle. Different letters above the columns indicate significant differences. Vehicle, the control rats which were orally gavaged with phosphate-buffered saline (PBS); CMTPTU, the rats that were colonized by cecal microbiota from PTU-treated donors; CMTYJT, the rats that were colonized by cecal microbiota from YJT + PTU-treated donors; CMTT4, the rats that were colonized by cecal microbiota from L-thyroxine+PTU -treated donors; CMTCon, the rats that were colonized with cecal microbiota from control donors.

Article Snippet: Published in partnership with Nanyang Technological University npj Biofilms and Microbiomes (2023) 32 Serum biochemical parameter assays We used enzyme-linked immunosorbent assay kits (ELISA) to estimate the levels in the serum, including TSH (CSB-E05115r), free Tri-iodothyronine (free-T3, CSB-E05076r), thyroxine (T4, CSBE05082r), GLP-1 (CSB-E08117r), TNF-α (CSB-E11987r), ghrelin (CSB-E09816r), and LPS (CSB-E14247r) following the manufacturer’s recommendations (Cusabio, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Control, Saline

Journal: NPJ biofilms and microbiomes

Article Title: Yijung-tang improves thermogenesis and reduces inflammation associated with gut microbiota in hypothyroid rats.

doi: 10.1038/s41522-023-00396-2

Figure Lengend Snippet: Fig. 7 The paradigm summarizing the thermogenic and anti-inflammatory properties of YJT via the gut microbiota and related multiple signaling pathways. Propylthiouracil (PTU)-treated hypothyroid rats exhibit hypothermia, hypophagia, and hyperinflammation, which have been associated with intestinal microbiota dysbiosis, particularly with extremely enriched Prevotellaceae. As a prebiotic, Yijung-tang (YJT) modulates the gut microbiota and metabolite-related receptors (e.g., FFAR3 and TGR5/FXR) to stimulate gene expression of intestinal tyrosine hydroxylase (Th) for norepinephrine synthesis and iodothyronine deiodinases 2 (Dio2) for thyroid hormone metabolism, and consequently activates the thermogenic pathway in brown adipose tissue (BAT) and attenuates hypothyroidism-related systemic inflammation. In contrast, L-thyroxine (T4) treatment in hypothyroid rats stimulates ghrelin secretion and activates intestinal LPS–TLR4 and peptidoglycan–Pglyrp2 signaling pathways, and leads to hyperthermia, hyperphagia, and hyperinflammation.

Article Snippet: Published in partnership with Nanyang Technological University npj Biofilms and Microbiomes (2023) 32 Serum biochemical parameter assays We used enzyme-linked immunosorbent assay kits (ELISA) to estimate the levels in the serum, including TSH (CSB-E05115r), free Tri-iodothyronine (free-T3, CSB-E05076r), thyroxine (T4, CSBE05082r), GLP-1 (CSB-E08117r), TNF-α (CSB-E11987r), ghrelin (CSB-E09816r), and LPS (CSB-E14247r) following the manufacturer’s recommendations (Cusabio, Wuhan, China).

Techniques: Protein-Protein interactions, Gene Expression