Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) MALDI-TOF spectra of reaction mixture. The ratio denotes the molar ratio of leptin monomer to polymer. (b) SEC of leptin conjugates eluted from Superdex75 100/300 GL column in 10% MeOH/0.25 M sodium phosphate, pH 7.5 at 0.5 ml/min. (c) SDS-PAGE of pooled fractions in SEC denoted by elution time. Fraction of 20.3–21.9 min in LepNP85 and fraction of 18.5–21 min in LepNPEG5K were used.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Polymer, SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: SDS-PAGE of the reaction mixture of leptin and Leptin-2PCA with CHO-P85-OH or with P85-DSP.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Leptin 1:1 physical mixture with P85 or PEG5K. (b) Leptin 1:1 conjugates.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Leptin and leptin 1:1 conjugates were eluted from Jupiter C4 column (particle diameter 5 μm, pore diameter 300 Å, 4.6 × 100 mm) by gradient elution at 1 ml/min and 25 °C, and monitored by absorption at 220 nm. Mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA: isopropanol 50:50 (v/v). The elution started from 5% B for 5 min, then linearly increased to 95% B at 1%/min, and stayed at 95% B till 100 min.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Representative sensorgram for native leptin. (b) Representative sensorgram for LepNP85 1:1 conjugates. (c) Representative sensorgram for LepDSSP85 1:1 conjugates. (d) Dissociation constants (KD) of leptin and leptin 1:1 conjugates. Data are mean ± SD, n= 5~12. *** p < 0.001 and n.s. not significant by One-way ANOVA and post Newman-Keuls Multiple Comparison Test.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Comparison

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Dose response of leptin and LepNP85 1:1 conjugates. Data are mean ± SEM, n=3. (b) Comparison of leptin and leptin 1:1 conjugates at 100 ng/mouse. Data are mean ± SEM, n= 5~7. ** p < 0.01 and *** p < 0.001 by One-way ANOVA and post Newman-Keuls Multiple Comparison Test.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Comparison

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Serum clearance, unidirectional brain influx rates and initial volumes of brain distribution for leptin and leptin 1:1 conjugates.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Male CD-1 mice were co-injected with 125I-LepNP85 and 131I-leptin. Data are mean ± SEM, n=7/time point, * p < 0.05, ** p < 0.01, and *** p < 0.001 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Each male CD-1 mouse was injected with 125I-labeled LepNP85 with or without 10 μg of cold leptin. Data are mean ± SEM, n=7/time point, * p < 0.05 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection, Labeling

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: 1 μg of biotin-labeled leptin and LepNP85 in 1 μl of PBS were injected locally into brain by ICV injection. The concentrations of biotin-labeled proteins in brain lysate and plasma were analyzed by ELISA. (a) Blood absorption of biotin-leptin and biotin-LepNP85 after ICV injection. (b) Brain retention of biotin-leptin and biotin-LepNP85 after ICV injection. Data are mean ± SEM, n=4/time point, * p < 0.05 by One-way ANOVA and post Dunnett’s Multiple Comparison Test, and *** p < 0.001 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Labeling, Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Comparison

Journal:

Article Title: The role of falling leptin levels in the neuroendocrine and metabolic adaptation to short-term starvation in healthy men

doi: 10.1172/JCI200317490

Figure Lengend Snippet: Pulsatility characteristics of leptin, LH, TSH, and GH on the third day of each fed or fasting state (n = 6)

Article Snippet: Conflict of interest: Christos S. Mantzoros has received research support and consulting fees from Amgen Inc. Nonstandard abbreviations used: recombinant-methionyl human leptin (r-metHuLeptin); General Clinical Research Center (GCRC); triiodothyronine (T3); sex hormone–binding globulin (SHBG); thyroxine (T4); free thyroxine (FT4); reverse T3 (rT3); thyroxine-binding globulin (TBG); IGF-binding proteins (IGFBPs); plasma renin activity (PRA); luteinizing hormone (LH); thyrotropin-stimulating hormone (TSH); gonadotrophin-releasing hormone (GnRH); thyrotropin-releasing hormone (TRH); dual energy X-ray absorptiometry (DEXA); area under the curve (AUC); hypothalamic-pituitary-adrenal (HPA); sympathetic nervous system (SNS); resting metabolic rate (RMR); hypothalamic-pituitary-gonadal (HPG); hypothalamic-pituitary-thyroid (HPT); leptin receptor (ObR); signal transducer and activator of transcription-3 (STAT3); extracellular signal-regulated kinase (ERK); neuropeptide Y (NPY).

Techniques:

Journal: Indian Journal of Endocrinology and Metabolism

Article Title: Leptin therapy, insulin sensitivity, and glucose homeostasis

doi: 10.4103/2230-8210.105571

Figure Lengend Snippet: Euglycemic hyperinsulinemic clamps of leptin-deficient patients while on leptin and after brief periods of leptin withdrawal (from Paz-Filho et al .). Glucose infusion rates increased in eight out of nine times when leptin therapy was briefly interrupted. The substantial weight gain after leptin withdrawal was responsible for acutely increasing insulin sensitivity

Article Snippet: [ ] Physiological doses of recombinant methionyl human leptin (r-metHuLeptin, Metreleptin ® , Amylin Pharmaceuticals, USA, 0.02-0.04 mg/kg/day) were initiated on the leptin-deficient individuals being assessed by our group at ages 5 (boy), 27 (male), 30 and 40 (females).

Techniques:

Journal: Indian Journal of Endocrinology and Metabolism

Article Title: Leptin therapy, insulin sensitivity, and glucose homeostasis

doi: 10.4103/2230-8210.105571

Figure Lengend Snippet: Interactions between fat, brain, pancreas and bone. Leptin activates the sympathetic tonus, which inhibits insulin secretion and stimulates Esp expression in the osteoblast (via stimulation of the adrenergic beta 2 receptor). Esp inhibits osteocalcin, decreasing insulin expression. Leptin inhibits pancreatic insulin. Insulin, in turn, stimulates leptin expression in the white adipose tissue. Adapted from Kieffer and Habener. H: Hypothalamus

Article Snippet: [ ] Physiological doses of recombinant methionyl human leptin (r-metHuLeptin, Metreleptin ® , Amylin Pharmaceuticals, USA, 0.02-0.04 mg/kg/day) were initiated on the leptin-deficient individuals being assessed by our group at ages 5 (boy), 27 (male), 30 and 40 (females).

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: The adipose-neural axis is involved in epicardial adipose tissue-related cardiac arrhythmias

doi: 10.1016/j.xcrm.2024.101559

Figure Lengend Snippet: Leptin secreted by adipocytes causes an irregular rhythm of cardiomyocytes by activating sympathetic neurons via the leptin receptor (A) Quantification of cytokines in the adipocyte supernatant using a bead-based multiplex assay ( n = 3 times per group). (B) Line-scan images of spontaneous Ca 2+ transients in hiPSC-CMs treated with IL-1β or IL-6 with or without hiPSC-SymNs. (C) Line-scan images of spontaneous Ca 2+ transients in hiPSC-CMs treated with IL-8 or TNF-α with or without hiPSC-SymNs. (D) Quantification of the beating rate and cells exhibiting irregular Ca 2+ transients in hiPSC-CMs cocultured with sympathetic neurons and a single cytokine ( n = 60 cells per group). Data are presented as the mean ± SEM. ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, determined by Student’s t test. (E) Line-scan images of spontaneous Ca 2+ transients and quantification of the beating rate and cells exhibiting irregular Ca 2+ transients in hiPSC-CMs treated with Lep with or without hipSC-SymNs ( n = 60 cells per group). Data are presented as the mean ± SEM. ∗∗∗ p < 0.001, determined by Student’s t test. (F) Line-scan images of spontaneous Ca 2+ transients and quantification of the beating rate and cells exhibiting irregular Ca 2+ transients in hiPSC-CMs treated with APN with or without hiPSC-SymNs ( n = 60 cells per group). Data are presented as the mean ± SEM. ns, not significant; ∗∗ p < 0.01, determined by Student’s t test. (G) Immunofluorescence staining for the sympathetic neuron marker TH and the leptin receptor (LepR). Scale bar, 50 μm. (H) Immunofluorescence assay for TH and the neuronal activity marker c-Fos in hiPSC-SymNs cultured with or without leptin. Scale bar, 50 μm. (I) The concentrations of neurotransmitters in the supernatant were analyzed using commercial ELISA kits before and after leptin administration ( n = 3 times per group). (J) Western blot analysis and relative quantification of STAT3, p-STAT3, and SOCS3 in hiPSC-SymNs treated with leptin, adi sup, or adi sup plus a JAK2 inhibitor (AG490) ( n = 3 times per group). (K) Line-scan images of spontaneous Ca 2+ transients and quantification of cells exhibiting irregular Ca 2+ transients in hiPSC-CMs from the triple coculture system with the addition of a leptin-neutralizing antibody ( n = 60 cells per group). Data are presented as the mean ± SEM. ∗ p < 0.05, ∗∗∗ p < 0.001, determined by one-way ANOVA test.

Article Snippet: Recombinant Human Leptin , PeproTech , Cat# AF-300-27-200.

Techniques: Multiplex Assay, Immunofluorescence, Staining, Marker, Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative Proteomics

Journal: Cell Reports Medicine

Article Title: The adipose-neural axis is involved in epicardial adipose tissue-related cardiac arrhythmias

doi: 10.1016/j.xcrm.2024.101559

Figure Lengend Snippet: Activation of sympathetic neurons leads to an irregular rhythm of cardiomyocytes via NPY/Y1R interactions (A) Line-scan images of spontaneous Ca 2+ transients in hiPSC-CMs from the triple coculture system treated with a blocker of α-adrenergic receptor, β-adrenergic receptor, or dopamine receptor 2. Red arrows indicate arrhythmia-like waveforms. (B) Quantification of the beating rate and cells exhibiting irregular Ca 2+ transients in hiPSC-CMs from the triple coculture system treated with a specific blocker ( n = 60 cells per group). Data are presented as the mean ± SEM. ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, determined by one-way ANOVA test. (C) qRT-PCR analysis of the gene expression of NPY receptors in hiPSC-CMs ( n = 3 times). (D) Line-scan images of spontaneous Ca 2+ transients and quantification of the beating rate and cells exhibiting irregular Ca 2+ transients in hiPSC-CMs from the triple coculture system treated with a Y1R blocker (BIBP3226) ( n = 60 cells per group). Data are presented as the mean ± SEM. ∗∗ p < 0.01, ∗∗∗ p < 0.001, determined by Student’s t test. (E) Line-scan images of spontaneous Ca 2+ transients and quantification of cells exhibiting irregular Ca 2+ transients in hiPSC-CMs treated with leptin, NPY or leptin plus NPY ( n = 60 cells per group). Data are presented as the mean ± SEM. ns, not significant; ∗∗∗ p < 0.001, determined by one-way ANOVA test.

Article Snippet: Recombinant Human Leptin , PeproTech , Cat# AF-300-27-200.

Techniques: Activation Assay, Quantitative RT-PCR, Gene Expression

Journal: Cell Reports Medicine

Article Title: The adipose-neural axis is involved in epicardial adipose tissue-related cardiac arrhythmias

doi: 10.1016/j.xcrm.2024.101559

Figure Lengend Snippet: Arrhythmogenesis is positively correlated with leptin and NPY levels in coronary sinus blood from human patients with AF (A) Leptin concentrations in CS blood and PV blood were detected and compared. (B) Correlations between the PV or CS leptin concentrations and EAT thickness. (C) The coronary sinus leptin concentrations were detected and compared in patients without AF ( n = 14), with paroxysmal AF ( n = 22), or with persistent AF ( n = 17). (D) Correlation between PV and CS NPY levels across all patients. (E) NPY concentrations in CS blood and PV blood were detected and compared ( n = 53). (F) Correlation between EAT thickness and the CS NPY concentration. (G) Correlation between CS leptin and CS NPY concentrations. (H) The CS NPY concentrations were compared between AF patients with different disease severities. (I) Line-scan images of spontaneous Ca 2+ transients and quantification of CMs from different groups of cells exhibiting irregular Ca 2+ transients ( n = 60 cells per group). Data are presented as the mean ± SEM. ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, determined by one-way ANOVA test.

Article Snippet: Recombinant Human Leptin , PeproTech , Cat# AF-300-27-200.

Techniques: Concentration Assay

Journal: Cell Reports Medicine

Article Title: The adipose-neural axis is involved in epicardial adipose tissue-related cardiac arrhythmias

doi: 10.1016/j.xcrm.2024.101559

Figure Lengend Snippet:

Article Snippet: Recombinant Human Leptin , PeproTech , Cat# AF-300-27-200.

Techniques: Recombinant, Clinical Proteomics, Derivative Assay, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Western Blot, Software

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Bone Mineral Content in Patients With Congenital Generalized Lipodystrophy Is Unaffected by Metreleptin Replacement Therapy

doi: 10.1210/jc.2014-1353

Figure Lengend Snippet: Baseline Characteristics

Article Snippet: The present study was an ancillary study of a protocol at the Clinical Center of the National Institutes of Health (NIH) designed to study the effects of recombinant human methionyl leptin (metreleptin; Bristol Myers Squibb) replacement on metabolic and hormonal derangements in patients with lipodystrophy ( {"type":"clinical-trial","attrs":{"text":"NCT00025883","term_id":"NCT00025883"}} NCT00025883 ).

Techniques: Mutagenesis