Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: European Journal of Nutrition

Article Title: Different infant formulas can activate toll-like receptor 9 in vitro and inhibit interleukin 6 in human primary intestinal epithelial cells

doi: 10.1007/s00394-024-03507-7

Figure Lengend Snippet: Expression and activity of TLR4 and TLR9 in human primary intestinal epithelial cells. a Western blot analysis of TLR4 and TLR9 in P-IECs, 0 h (basal) and 24 h after stimulation with TLR4 (LPS) or TLR9 agonist (ODN2006). GAPDH was used as reference protein. b Quantitative RT-PCR showing the levels of TLR4 and TLR9 in P-IECs 0 h, 2 h, and 6 h after stimulation with TLR4 (LPS) or TLR9 agonist (ODN2006). GAPDH was used as reference gene. n = 7/group. c, d HEK-Dual cells were stimulated with TLR9 agonist (ODN2006) for 2 h, 6 h, 11 h, and 24 h and the ( c ) optical density (OD 630 ) or ( d ) relative light units (RLUs) were compared with non-treated cells (basal). n = 4/group. e, f HEK-Dual cells were treated with a TLR9 agonist (A, ODN2006) or with inhibitor (INH-18) and agonist (I + A) in two different concentrations, 0.1 µg/ml ( e ) and 1.0 µg/ml ( f ), and were compared to non-treated cells. Luminescence signal was measured with a TECAN luminescence reader. n = 4/group. g MTT assay showing cell viability in different dilutions of the control formula, compared to control (Co) cells incubated with medium. n = 8/group. Data were analyzed by two-way ANOVA ( b–d ) or one-way ANOVA ( g ) followed by Newman-Keuls post hoc test or Student’s t -test ( e, f ) and are represented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Adult human primary intestinal epithelial cells (P-IECs) were purchased from PELOBiotech GmbH (Martinsried, Germany).

Techniques: Expressing, Activity Assay, Western Blot, Quantitative RT-PCR, MTT Assay, Control, Incubation

Journal: European Journal of Nutrition

Article Title: Different infant formulas can activate toll-like receptor 9 in vitro and inhibit interleukin 6 in human primary intestinal epithelial cells

doi: 10.1007/s00394-024-03507-7

Figure Lengend Snippet: IL6 levels were decreased through various infant formulas in human primary intestinal epithelial cells; in contrast IL6 levels were increased due to TLR9 inhibition. The following phosphorylation stages and proteins were investigated by Western blot analysis in human P-IECs: TLR9, TLR4, pMAPK1/2 and MAPK1/2, pMAPK8/9 and MAPK8/9, pp38 and p38, pIRAK4 and IRAK4, IL6, NFKB, pIKB and IKB. HEK-Dual cells were cultivated in medium, PF, CF, and PHF in a 1/25 dilution, with TLR9 and TLR4 inhibitor (0.1 µg/ml) and non-treated HEK-Dual cells served as reference. In addition, the cells were incubated with L. fermentum CECT5716 (LC) (10 10 CFUs) for 30 min. GAPDH was used as reference protein

Article Snippet: Adult human primary intestinal epithelial cells (P-IECs) were purchased from PELOBiotech GmbH (Martinsried, Germany).

Techniques: Inhibition, Western Blot, Incubation