Journal: Metabolic Brain Disease

Article Title: The SIRT-1/Nrf-2/HO-1 antioxidant defense axis in adult attention-deficit/hyperactivity disorder

doi: 10.1007/s11011-026-01845-5

Figure Lengend Snippet: ROC Curve Analysis for HO-1, Nrf-2 and SIRT-1. HO-1 (cut-off value 0.87, sensitivity 78.3%, specificity 76.7%); Nrf-2 (cut-off value 9.8 sensitivity 70%, specificity 73.3%); SIRT-1 (cut-off value 8.3, sensitivity 75%, specificity 70%). Abbreviations: HO-1: heme oxygenases-1; Nrf-2: nuclear factor erythroid 2-related factor 2; SIRT-1: Sirtuin 1

Article Snippet: Serum HO-1, NRF-2, and SIRT-1 concentrations were analyzed using ELISA kits according to the manufacturers’ standard protocols (BT Lab, Human Heme Oxygenase-1: Cat. No. E0932Hu; BT Lab, Human Nuclear Factor Erythroid 2-Related Factor 2: Cat. No. E3244Hu; BT Lab, Human Sirtuin-1: Cat. No. E2557Hu; Jiaxing Korain Biotech, Jiaxing, China) on a Rel Assay automated ELISA reader (Biobase Biodusty Co., Ltd., Jinan, China).

Techniques:

Journal: F1000Research

Article Title: Exploring the moderating effects of SIRT1 and gene polymorphisms rs7895833 on the relationship between hemoglobin levels and physical frailty in elderly adults with comorbid chronic diseases: A moderated mediation analysis

doi: 10.12688/f1000research.133517.3

Figure Lengend Snippet: Figure 1. SIRT1 SNP rs7895833 A>G representative PCR gel electrophoresis images.

Article Snippet: The plasma levels of SIRT1 was determined by a monoclonal antibody-based ELISA method using a commercially available human SIRT1 ELISA kit (Elabscience®, Houston, USA, E-EL-H1546).

Techniques: Nucleic Acid Electrophoresis

Journal: F1000Research

Article Title: Exploring the moderating effects of SIRT1 and gene polymorphisms rs7895833 on the relationship between hemoglobin levels and physical frailty in elderly adults with comorbid chronic diseases: A moderated mediation analysis

doi: 10.12688/f1000research.133517.3

Figure Lengend Snippet: Figure 2. The correlation plot of the interrelationship between the potential main variables. Hb (Hemoglobin); SIRT1 (Sirtuin1); Frail score (frailty); genotype (AA, AG-GG). Spearman's rank correlation with two tail significant: *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The plasma levels of SIRT1 was determined by a monoclonal antibody-based ELISA method using a commercially available human SIRT1 ELISA kit (Elabscience®, Houston, USA, E-EL-H1546).

Techniques:

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 treatment reduces plasma LDL-cholesterol and protects against atherosclerosis in ApoE −/− mice. Eight-week-old ApoE −/− mice were fed on a high cholesterol diet (1.25% w/w) for 4 weeks and randomized to be treated with rmSIRT1 ( n = 6) or vehicle (PBS containing 0.1% BSA) ( n = 6) for another 4 weeks. ( A ) Plasma concentration of SIRT1 measured using ELISA of C57BL/6J wild-type mice ( n = 6), ApoE −/− mice + vehicle ( n = 6) and ApoE −/− mice + rmSIRT1 ( n = 6). ( B ) Weekly measurements of body weight of ApoE −/− mice + vehicle ( n = 6) and ApoE −/− mice + rmSIRT1 ( n = 6). ( C ) Graph showing cholesterol distribution in the different lipoprotein sub-fractions separated by gel filtration chromatography; ( D ) Bar graph of plasma total cholesterol and ( E ) LDL-cholesterol concentrations. ( F ) Representative pictures (left) and quantifications of ( G ) thoracic-abdominal aortae en face , ( H ) aortic root cross sections stained with ORO and ( I ) immunohistochemically for macrophages (CD68). Scale bars in photomicrographs: 1 mm (for E ) and 500 μm (for F , G ). Grey bars represent vehicle treatment and green bars represent rmSIRT1 treatment. ORO, Oil-Red O. Values are represented as means ± SEM. Statistical significance was performed using Student’s unpaired t -test.

Article Snippet: Where indicated, cells were treated with vehicle control (Phosphate buffer saline containing 0.1% BSA) or recombinant human SIRT1 (rhSIRT1, 1 μmol/L) (CSB-EP822202HU, Cusabio Biotech, LubioScience, Switzerland) or recombinant human PCSK9 (rhPCSK9, ab198471, Abcam, concentrations in Figure legends) for 2 h.

Techniques: Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay, Filtration, Chromatography, Staining

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 increases hepatic LDLR by post-translational modification of PCSK9 in ApoE −/− mice. Eight-week-old ApoE −/− mice were fed on a high cholesterol diet (1.25% w/w) for 4 weeks and were randomized to be treated with rmSIRT1 ( n = 6) or with vehicle (PBS with 0.1% BSA) ( n = 6) for another 4 weeks. ( A ) Western blot of LDLR (120 kDa) and PCSK9 (∼62 kDa) in hepatic tissue lysates of mice ( n = 5). β-actin (45 kDa) was used as loading control. Bar graph of ELISA of ( B ) total PCSK9 and ( C ) acetylated PCSK9 in mice plasma. Statistical significance was performed using Student’s unpaired two-sample t -test. ( D , E , F ) Human hepatoma Huh7 cells [three independent triplicate experiments ( n = 3)] were treated with either 1 μmol/L of recombinant human SIRT1 (rhSIRT1) or vehicle (PBS containing 0.1% BSA) for 2 h at 37°C. The cell culture media was collected, and cells were lysed to perform ELISA to detect PCSK9. Bar graph of ELISA of ( D ) total cellular PCSK9, ( E ) total media PCSK9 and ( F ) acetylated PCSK9 in the media of cultured Huh7 cells. Grey bars represent vehicle treatment and green bars represent rmSIRT1 or rhSIRT1 treatment as indicated. Data are represented as means ± SEM. Statistical significance was performed using Student’s unpaired t -test.

Article Snippet: Where indicated, cells were treated with vehicle control (Phosphate buffer saline containing 0.1% BSA) or recombinant human SIRT1 (rhSIRT1, 1 μmol/L) (CSB-EP822202HU, Cusabio Biotech, LubioScience, Switzerland) or recombinant human PCSK9 (rhPCSK9, ab198471, Abcam, concentrations in Figure legends) for 2 h.

Techniques: Modification, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Recombinant, Cell Culture

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 inhibits PCSK9-mediated LDLR degradation in hepatocytes. HuH-7 (hereafter Huh7) cells were transfected with a specific siRNA against LDLR or with non-silencing control siRNA (NS control). Assays were performed 72 h post transfection. Cells were then pre-treated with 1 μmol/L of rhSIRT1 for 2 h ( A ) Specific cellular binding of 125 I-LDL was measured at 4°C ( B ) Specific cellular association of 125 I-LDL was measured at 37°C. ( C , D , E ) Huh7 cells were co-incubated with rhPCSK9 (2 μg/mL) with rhSIRT1 (1 μmol/L) for 2 h at 37°C ( C ) Representative Western blots ( n = 3) and quantification of blot density of LDLR (130 kDa) degradation in Huh7 cells. Vinculin (140 kDa) was used as the loading control. ( D ) Specific cellular binding of 125 I-LDL was measured at 4°C ( E ) Specific cellular association of 125 I-LDL was measured at 37°C. Values are represented as means ± SEM of three independent triplicate or more experiments ( n = 3). Statistical significance was performed using one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: Where indicated, cells were treated with vehicle control (Phosphate buffer saline containing 0.1% BSA) or recombinant human SIRT1 (rhSIRT1, 1 μmol/L) (CSB-EP822202HU, Cusabio Biotech, LubioScience, Switzerland) or recombinant human PCSK9 (rhPCSK9, ab198471, Abcam, concentrations in Figure legends) for 2 h.

Techniques: Transfection, Control, Binding Assay, Incubation, Western Blot, Comparison

Journal: Cardiovascular Research

Article Title: Sirtuin-1 directly binds and deacetylates hepatic PCSK9 thereby promoting the inhibition of LDL receptor degradation

doi: 10.1093/cvr/cvaf087

Figure Lengend Snippet: Exogenous SIRT1 inhibits PCSK9 activity through direct deacetylation in hepatocytes—specific mutation of these deacetylation sites restores PCSK9 activity. Huh7 cells were incubated with rhSIRT1 (1 μmol/L) for 2 h at 37°C. ( A ) Western blot analysis of PCSK9 immunoprecipitated Huh7 cells lysates with total PCSK9 antibody (∼62 kDa) and acetyl lysine antibody (Ac-K-PCSK9, ∼62 kDa). ( B ) Direct binding assay interaction between rhSIRT1 and rhPCSK9 using Surface Plasmon Resonance. The equilibrium dissociation constant was 34 nM. ( C ) Schematic diagram of PCSK9 domains and an overview of lysine(K) sites on PCSK9 deacetylated by SIRT1. Representative Western blot and quantification of LDLR (130 kDa) and Vinculin (140 kDa) expression in Huh7 cells expressing WT, 3KR, 3KQ mutants of PCSK9 for 72 h ( n = 3). (E )V. means empty vector ( D ) Specific cellular binding of 125 I-LDL was measured at 4°C ( E ) Specific cellular association of 125 I-LDL was measured at 37°C. ( F ) Binding of acetylated and deacetylated PCSK9 to EGF-A of LDLR using ELISA. SP—signal peptide, Pro—pro-domain, Catalytic—catalytic domain, CHRD—cysteine, histidine-rich domain. Values are represented as means ± SEM of three independent triplicate or more experiments ( n = 3). Statistical significance was performed using one-way ANOVA followed by Tukey’s multiple comparison test.

Article Snippet: Where indicated, cells were treated with vehicle control (Phosphate buffer saline containing 0.1% BSA) or recombinant human SIRT1 (rhSIRT1, 1 μmol/L) (CSB-EP822202HU, Cusabio Biotech, LubioScience, Switzerland) or recombinant human PCSK9 (rhPCSK9, ab198471, Abcam, concentrations in Figure legends) for 2 h.

Techniques: Activity Assay, Mutagenesis, Incubation, Western Blot, Immunoprecipitation, Binding Assay, SPR Assay, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Aging (Albany NY)

Article Title: Scavengers of reactive γ-ketoaldehydes extend Caenorhabditis elegans lifespan and healthspan through protein-level interactions with SIR-2.1 and ETS-7

doi: 10.18632/aging.101011

Figure Lengend Snippet: ( A ) Synthetically purified IsoKs decrease biochemical activity of rhSIRT1. Recombinant human SIRT1 was incubated with increasing concentrations of IsoK and enzymatic activity was assessed using a luminescence based assay. Concentration-response curves were generated and IC 50 values were calculated from three independent experiments. ( B ) Kaplan-Meier survival curves depicting effects of SA administration on lifespan of non-functional SIR-2.1 mutant. ( C ) Summary of SA-treated SIR-2.1 mutant median lifespan. SA administration does not affect median lifespan of SIR-2.1 mutants. Data are expressed as means ± SEM from four independent experiments. P = 0.70. ( D ) Changes in lipofuscin autofluorescence accumulation with age. Compared to vehicle control in WT animals, SA response profiles indicate neither dose of SA were able to decrease the accumulation of lipofuscin. Data are expressed as means ± SEM from four independent experiments. P = 0.5. ( E ) Changes in pharyngeal pumping rate in SA-treated SIR-2.1 mutants. Administration of SA failed to preserve pharyngeal pumping rate. Data are expressed as means ± SEM from four independent experiments. P = 0.5.

Article Snippet: Purified recombinant human SIRT1 (R&D Systems, Biotechne) activity was assayed in HEPES-buffered saline (10 mM HEPES, 150 nM NaCl, 2 mM MgCl2) in the presence and absence of 15-E 2 -IsoK.

Techniques: Purification, Activity Assay, Recombinant, Incubation, Luminescence Assay, Generated, Functional Assay, Mutagenesis

Journal: Antioxidants

Article Title: The Effect of Repeated Whole-Body Cryotherapy on Sirt1 and Sirt3 Concentrations and Oxidative Status in Older and Young Men Performing Different Levels of Physical Activity

doi: 10.3390/antiox10010037

Figure Lengend Snippet: Level of sirtuins and markers of oxidative stress in older male before and after the application of whole-body cryotherapy (WBC).

Article Snippet: The concentration of sirtuins was determined in the serum via the enzyme immunoassay (ELISA) method using the high-sensitivity Sirt1 SEE912Hu and Sirt3 SEE913Hu reagent kits (Cloud—Clone Corp., Houston, TX, USA).

Techniques:

Journal: Antioxidants

Article Title: The Effect of Repeated Whole-Body Cryotherapy on Sirt1 and Sirt3 Concentrations and Oxidative Status in Older and Young Men Performing Different Levels of Physical Activity

doi: 10.3390/antiox10010037

Figure Lengend Snippet: Level of sirtuins and markers of oxidative stress in young males before and after the application of whole-body cryotherapy (WBC).

Article Snippet: The concentration of sirtuins was determined in the serum via the enzyme immunoassay (ELISA) method using the high-sensitivity Sirt1 SEE912Hu and Sirt3 SEE913Hu reagent kits (Cloud—Clone Corp., Houston, TX, USA).

Techniques:

Journal: Antioxidants

Article Title: The Effect of Repeated Whole-Body Cryotherapy on Sirt1 and Sirt3 Concentrations and Oxidative Status in Older and Young Men Performing Different Levels of Physical Activity

doi: 10.3390/antiox10010037

Figure Lengend Snippet: Correlations between the concentrations of sirtuins and prooxidative–antioxidant balance markers in older training and non-training men.

Article Snippet: The concentration of sirtuins was determined in the serum via the enzyme immunoassay (ELISA) method using the high-sensitivity Sirt1 SEE912Hu and Sirt3 SEE913Hu reagent kits (Cloud—Clone Corp., Houston, TX, USA).

Techniques:

Journal: Antioxidants

Article Title: The Effect of Repeated Whole-Body Cryotherapy on Sirt1 and Sirt3 Concentrations and Oxidative Status in Older and Young Men Performing Different Levels of Physical Activity

doi: 10.3390/antiox10010037

Figure Lengend Snippet: Correlations between the concentrations of sirtuins and prooxidative–antioxidant balance markers in young training and non-training men.

Article Snippet: The concentration of sirtuins was determined in the serum via the enzyme immunoassay (ELISA) method using the high-sensitivity Sirt1 SEE912Hu and Sirt3 SEE913Hu reagent kits (Cloud—Clone Corp., Houston, TX, USA).

Techniques: