Journal: The Journal of Biological Chemistry

Article Title: Loss of Phosphatase and Tensin Homolog (PTEN) Induces Leptin-mediated Leptin Gene Expression

doi: 10.1074/jbc.M113.481523

Figure Lengend Snippet: H1650 human lung adenocarcinoma cells are PTEN-deficient and express high leptin and leptin receptor mRNA. Comparison of different lung cancer cell lines reveals H1650 cells are PTEN-deficient (A) and express the highest levels of LEP and its receptor LEPR mRNA (B). C, leptin and leptin receptor protein levels as a result of PTEN deletion as assessed in H1650 cells, suggesting the likely presence of a functional LEP signaling pathway in lung epithelial cells. D, dose-dependent increase in H1650 cell proliferation (∼3-fold) was observed when cells were treated for 48 h with increasing concentrations (50, 100, and 200 ng/ml; lane 2–4) of human recombinant LEP. E, continuous impedance sensing measurements identify greater wound closure efficiency in cells treated with 100 ng/ml leptin prior to wounding as compared with control cells without leptin. F, identification of the LEP gene core promoter region in H1650 lung cancer cell line revealed a proximal enhancer containing NRF-1 and δ-binding sites. Subconfluent cultures were transiently transfected with various promoter-reporter deletion constructs derived from 5′-upstream regulatory sequence of the LEP gene. Luciferase activity was expressed relative to the base-line luciferase activity of a promoter-less luciferase reporter construct (pGL3-Basic) set to unity. Data are represented from four independent experiments performed in triplicate (± S.E.; *, p value ≤ 0.05). G, diagrammatic representation of the most active promoter region −149 to +21 bp of the LEP gene (Luc-150), including proximal enhancers depicting the positions of NRF-1 and C/EBP sites.

Article Snippet: H1650 cells were grown on electric cell substrate impedance sensing 8-well plate arrays (8W1E; Applied Biophysics, Troy, NY) in growth media with serum until fully confluent, after which the media were replaced with serum-free media for 24 h. Serum-deprived cells were treated with 100 ng/ml of human recombinant LEP (R&D Systems, Minneapolis, MN) for 2 h prior to wounding.

Techniques: Comparison, Functional Assay, Recombinant, Binding Assay, Transfection, Construct, Derivative Assay, Sequencing, Luciferase, Activity Assay

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: 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