Journal: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison

Journal: EBioMedicine

Article Title: Ketogenesis acts as an endogenous protective programme to restrain inflammatory macrophage activation during acute pancreatitis

doi: 10.1016/j.ebiom.2022.103959

Figure Lengend Snippet: βOHB limits proinflammatory macrophage activation via class I HDACs, (a, b) Gene set enrichment analysis (GSEA) of gene expression in murine BMDMs are depicted. (a) Enrichment of TSA-upregulated signature genes (GSE22049) in βOHB-treated BMDMs. (b) Enrichment of HDAC3 depletion related signature genes (GES33162) in βOHB-treated BMDMs. (c, d) BMDMs treated with LPS (100 ng/ml) together with βOHB (10 mM), TSA (1 μΜ), or MS-275 (20 μM) for 4 h. (c) Flow cytometry analysis of intracellular TNFα levels. (d) Quantitative PCR analysis of Nos2, Il6, Il12b , and Tnfa mRNA expression. (e) ChIP-qPCR analysis of H3Ac and H4Ac occupancy in Edn1, Il12a , and Nos2 loci in the indicated group, with IgG as the control. (f) Ingenuity Pathway Analysis (IPA) of genes in cluster A. (g) BMDMs were pretreated with βOHB (10 mM) or TSA (1 μΜ) 1 h prior to LPS (100 ng/ml) stimulation for 15 min. Acylated p65 levels were determined by immunoprecipitation, while p-p65 and total p65 were determined by immunoblotting. (h) Mice were i.p. injected with TSA (0.1 mg/kg), MS-275 (20 mg/kg) or DMSO as a control 1 h prior to the CER-AP (Cer*7) induction and were harvested at 24 h thereafter (n = 5 / group). Frequency of TNFα + macrophagess (F4/80 + CD11b + ) in pancreatic leukocytes from the indicated groups. Data are presented as the mean ± SD in bar graphs (c–e) or the mean ± SEM in dot plots (h). *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: For treatments, the Balb/c mice were (1) fed with 1,3-butanediol (5.72% (w/v); Sigma-Aldrich) or normal water (control) one week before disease induction; received a single intraperitoneal injection of βOHB (3 mmol/kg; Sigma-Aldrich) or normal saline at the indicated time; and (3) the CPT1α inhibitor etomoxir (20 mg/kg, Sigma-Aldrich), histone deacetylase (HDAC) pan inhibitor TSA (0.1 mg/kg; Targetmol), HDAC1/3 inhibitor MS-275 (20 mg/kg; Targetmol), or dimethyl sulfoxide (DMSO) as a control 1 h before disease induction.

Techniques: Activation Assay, Gene Expression, Flow Cytometry, Real-time Polymerase Chain Reaction, Expressing, ChIP-qPCR, Control, Immunoprecipitation, Western Blot, Injection

Journal: Cancer Science

Article Title: Fragmentation of cell‐free DNA is induced by upper‐tract urothelial carcinoma–associated systemic inflammation

doi: 10.1111/cas.14679

Figure Lengend Snippet: A, The comparison of the concentration of each cytokine between the group with shorter plasma cell‐free DNA (cfDNA) fragment size (<166 bp) and the group with longer plasma cfDNA fragment size (166 bp≤). A comparison between the two groups was performed by Wilcoxon test, * P < .05, ** P < .01. B, The scatterplot of the fragment size of plasma cfDNA and the serum level of each cytokine (tumor necrosis factor‐α [TNFα], interleukin‐6 [IL‐6], interleukin‐1 receptor antagonist [IL‐1ra]) in upper‐tract urothelial carcinoma (UTUC) patients. The blue line shows the correlation curve, and the grey area represents a 95% confidence interval [CI]

Article Snippet: PBMCs cultures were incubated with the three cytokines as follows: TNFα (1‐100 ng/mL), IL‐6 (10‐100 ng/mL), or IL‐1ra (10‐100 ng/mL) (FUJIFILM).

Techniques: Concentration Assay

Journal: Cancer Science

Article Title: Fragmentation of cell‐free DNA is induced by upper‐tract urothelial carcinoma–associated systemic inflammation

doi: 10.1111/cas.14679

Figure Lengend Snippet: Measurement of the proportion of short fragment fraction in cell‐free DNA (cfDNA) derived from peripheral blood mononuclear cells [PBMCs]. Proportions of short fragment fraction of cfDNA in the culture supernatant of cytokine‐untreated PBMCs and TNFα‐treated PBMCs (10 ng/mL, 100 ng/mL). (* P < .05). A, Proportions of short fragment fraction of cfDNA in the culture supernatant of cytokine‐untreated PBMCs and interleukin‐6 (IL‐6)‐treated PBMCs (10 ng/mL, 100 ng/mL). B, Proportions of short fragment fraction of cfDNA in the culture supernatant of cytokine‐untreated PBMCs and IL‐1ra–treated PBMCs (10 ng/mL, 100 ng/mL). A comparison among the three groups was performed by Dunn's multiple comparison test, * P < .05

Article Snippet: PBMCs cultures were incubated with the three cytokines as follows: TNFα (1‐100 ng/mL), IL‐6 (10‐100 ng/mL), or IL‐1ra (10‐100 ng/mL) (FUJIFILM).

Techniques: Derivative Assay