Journal: BMB Reports

Article Title: Effects of α-lipoic acid on LPS-induced neuroinflammation and NLRP3 inflammasome activation through the regulation of BV-2 microglial cells activation

doi: 10.5483/BMBRep.2019.52.10.026

Figure Lengend Snippet: α-LA inhibits the expression of pro-inflammatory cytokines in LPS-treated BV-2 microglial cells. (A) Effects of α-LA on cell viability. BV-2 microglial cells were incubated with LPS (1 μg/ml) for 30 min followed by treatment with the indicated concentrations of α-LA for 24 h. Thereafter, cell viability was assessed through the MTT assay. (B and C) BV-2 microglial cells were treated with LPS (1 μg/ml) for 30 min followed by treatment with the indicated concentrations of α-LA at the suggested times. The cell-free conditioned culture medium was collected and analyzed with ELISA for TNF-α, IL-6. Data from three independent experiments are presented as means ± S.D. *≤ 0.05, **< 0.01, ***< 0.001 and are related to both LPS-induced cells and α-LA treated cells.

Article Snippet: Both TNF-α and IL-6 were quantitatively measured through an enzyme-linked immunosorbent assay (ELISA) using the mouse TNF-α and IL-6 DuoSet ELISA kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Incubation, MTT Assay, Enzyme-linked Immunosorbent Assay

Journal: JCI Insight

Article Title: Quinolinic acid potentially links kidney injury to brain toxicity

doi: 10.1172/jci.insight.180229

Figure Lengend Snippet: Kidney cortex RNA and protein lysates were prepared from Mdm2 -cKO vs. control mice ( n = 4 per group). ( A ) Transformation related protein 53 ( Trp53 ) mRNA and ( B ) p53 protein levels. ( C ) Cyclin-dependent kinase inhibitor 1A ( Cdkn1a ) mRNA and ( D ) p21 protein levels. ( E ) Cytochrome b-245, beta polypeptide ( Cybb ), mRNA and ( F ) CYBB protein levels. ( G ) CXC chemokine ligand 1 ( Cxcl1 ) mRNA level. ( H ) CXCL1 protein concentration. ( I ) Interleukin-1β ( Il1b ) mRNA level. ( J ) IL-1β protein concentration. ( K ) Neutrophil-to-lymphocyte ratio (VetScan Analyzer). ( L ) Plasma interleukin-6 (IL-6) protein concentration. ( M ) Plasma soluble urokinase plasminogen activator receptor (suPAR) concentration. Quantitative PCR (qPCR) results (normalized to Gapdh expression) and protein levels measured via Western blot (normalized to actin) are presented as fold-change relative to control. The proteins analyzed via ELISA are reported as concentrations. Graphs display means ± SEM. Two-tailed t tests: * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Quantitative protein levels in plasma and tissue lysates were assessed using ELISA kits following manufacturer’s protocols: mouse uPAR DuoSet ELISA (R&D Systems, Bio-Techne, DY531), mouse CXCL1/KC Quantikine ELISA Kit (R&D Systems, Bio-Techne, MKC00B-1), mouse IL-6 Quantikine ELISA Kit (R&D Systems, Bio-Techne, M6000B-1), and mouse IL-1β/IL-1F2 Quantikine ELISA Kit (R&D Systems, Bio-Techne, MLB00C-1).

Techniques: Control, Transformation Assay, Protein Concentration, Clinical Proteomics, Concentration Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: JCI Insight

Article Title: Quinolinic acid potentially links kidney injury to brain toxicity

doi: 10.1172/jci.insight.180229

Figure Lengend Snippet: Mice were fed regular chow (control) or 0.2% adenine-enriched chow for 1 month ( n = 4 per group). ( A and B ) Plasma suPAR and IL-6 levels measured by ELISA. Concentrations of Trp, KYN, 3HK, QA, and 5-HIAA in ( C – G ) plasma, kidney cortex ( H – L ), and brain cortex ( M – Q ). QA and 5-HIAA were measured using a different mass spectrometry method. Graphs display means ± SEM. Two-tailed t test; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Quantitative protein levels in plasma and tissue lysates were assessed using ELISA kits following manufacturer’s protocols: mouse uPAR DuoSet ELISA (R&D Systems, Bio-Techne, DY531), mouse CXCL1/KC Quantikine ELISA Kit (R&D Systems, Bio-Techne, MKC00B-1), mouse IL-6 Quantikine ELISA Kit (R&D Systems, Bio-Techne, M6000B-1), and mouse IL-1β/IL-1F2 Quantikine ELISA Kit (R&D Systems, Bio-Techne, MLB00C-1).

Techniques: Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Mass Spectrometry, Two Tailed Test

Journal: Clinical Cancer Research

Article Title: IFNα Augments Clinical Efficacy of Regulatory T-cell Depletion with Denileukin Diftitox in Ovarian Cancer

doi: 10.1158/1078-0432.ccr-20-4594

Figure Lengend Snippet: Figure 5. DCs mediate IFNa effects on Treg suppression through IL6. A, Flow cytometric analysis of suppression of T-cell proliferation by Tregs from the PEC of ID8- challenged IFNR/ mice adoptively transferred with WT or IFNR/ DCs and then treated with DD plus IFNa (samples pooled for each experiment, all groups n ¼ 3). B, Luminex analysis of IL6 and IL10 in supernatant with Tregs, DCs and/or IFNa (one of three experiments with similar results). Suppression of T-cell proliferation by Tregs in the presence of DCs, IFNa, and/or anti-IL6 (n ¼ 3 for all groups; C) or in the presence of DCs, IFNa, and/or recombinant IL6 (n ¼ 3 for all groups; D). E, Suppression of T-cell proliferation by Tregs in the presence of DCs, recombinant IL6, anti-CD80 and/or anti-CD86 (n ¼ 3 for all groups). F, Treg function in mLNs of mice challenged with ID8, treated with DDþIFNa PBS or anti-IL6 in vivo (n ¼ 5 for both groups).

Article Snippet: Recombinant mouse IL6 (R&D Systems) at 0.1 or 1.0 mg/mL was added withoutDCs as indicated.

Techniques: Luminex, Recombinant, In Vivo

Journal: Clinical Cancer Research

Article Title: IFNα Augments Clinical Efficacy of Regulatory T-cell Depletion with Denileukin Diftitox in Ovarian Cancer

doi: 10.1158/1078-0432.ccr-20-4594

Figure Lengend Snippet: Figure 7. Pegylated IFNa has activity in the clinical response to failed DD in patients with ovarian cancer. A, CA-125 serum levels in patient 103. IFNa given as weekly, subcutaneous pegylated IFNa2a. B, CA-125 serum levels in patient 105. C, Serum IL6 in patient 103 after treatment with DD alone and after addition of pegylated IFNa2a. Mean of triplicate replicates. D, Flow cytometric analysis of Treg prevalence (CD4þCD25hiCD127lo cells among all blood CD3þCD4þ

Article Snippet: Recombinant mouse IL6 (R&D Systems) at 0.1 or 1.0 mg/mL was added withoutDCs as indicated.

Techniques: Activity Assay

Journal: ADMET & DMPK

Article Title: Anti-inflammatory potential of plant-derived extracellular vesicles from Solanum nigrum L. integrated in gelatine-dopamine hydrogel on RAW 264.7 and MC3T3 cells

doi: 10.5599/admet.3149

Figure Lengend Snippet: Interleukin-6 (IL-6) concentration by (A) RAW 264.7 and (B) MC3T3 cells LPS induction and PDEV (2.5, 5 and 10 μg mL -1 ) treatment released from Gel-Dop hydrogel. Data presented as mean ± standard deviation (*** p < 0.001, **** p <0,0001)

Article Snippet: Other materials used in this study were Dulbecco’s modified eagle medium (DMEM) - high glucose, foetal bovine serum (FBS), Antibiotic-Antimycotic (ABAM), polyethylene glycol (PEG) 6000, trehalose (Sigma-Aldrich; Merck), phosphate-buffered saline (PBS), sodium periodate (NaIO 4 ) (Sigma-Aldrich), bovine serum albumin (BSA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT), 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), paraformaldehyde (PFA), kit BCA Assay (PierceTM BCA Protein Assay Kit, Thermo Scientific), PKH67 Green Fluorescent Cell Linker Kits (Sigma-Aldrich; Merck), and Mouse Interleukin-6 (IL-6) ELISA Kit (BT Lab, Cat. No. E0049Mo).

Techniques: Concentration Assay, Standard Deviation