Journal: PLOS Neglected Tropical Diseases

Article Title: In vitro and in vivo endothelial interactions of Leptospira species are markers of virulence

doi: 10.1371/journal.pntd.0013939

Figure Lengend Snippet: (top) HMEC-1 cells were grown to post-confluence, bacteria were added at an MOI of 20 and incubated for one hour. Non-associated bacteria were removed by washing and qPCR was performed to quantify associated bacteria. Strains associate with endothelial cells to varying extents, with 3/6 P1 + strains and 2/5 P1- species binding significantly more than Lb P. (bottom) HMEC-1 cells were grown to confluence on glass coverslips and infected at an MOI of 20 for 24 h. After washing to remove unbound bacteria, cells were fixed, stained for VE-cadherin, and mounted with Prolong Diamond Antifade Mountant with DAPI. Binary area was quantified as previously described and subtracted from uninfected controls to define “VE-cadherin disruption”. P1 + Leptospira (4/6) disrupt VE-cadherin more than other clades (3/10). Spearman correlation analysis determines cell association and VE-cadherin disruption correlate with r s = 0.644 and p = 0.0085. Strains are ordered based upon presence of virulence-associated genes in their genomes . Mean ± SEM is plotted. Each column is compared to LbP . * p < 0.05, # p < 0.01, & p < 0.001, $ p < 0.0001.

Article Snippet: Human dermal microvascular endothelial cells (HMEC-1) [ ] were obtained from ATCC (CRL-3243) and cultured as described previously [ , ] in MCDB medium at 37°C with 5% CO 2 .

Techniques: Bacteria, Incubation, Binding Assay, Infection, Staining, Disruption

Journal: Cells

Article Title: The Role of Aldosterone in Vascular Permeability in Diabetes

doi: 10.3390/cells15010089

Figure Lengend Snippet: Permeability of the human dermal microvascular endothelial cell (HMEC-1) monolayer under normal (NORM) or hyperglycemic (HG) conditions after MR blockade; ( a ) with 15 min of aldosterone exposure, and ( b ) with 60 min of aldosterone exposure. ALDO—aldosterone; EPL—eplerenone. Results are presented as mean ± SD; n = 6–9. Statistical relationships among groups were visualized using compact letter displays, where groups sharing the same letter are not significantly different, whereas groups without a common letter differ significantly ( p < 0.05).

Article Snippet: Immortalized human dermal microvascular endothelial cells (HMEC-1; ATCC ® CRL-3243TM; ATCC, Glasgow, UK) were maintained at 37 °C in a 5% CO 2 atmosphere in MCB131 medium (GibcoTM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum, streptomycin–penicillin (100 U/mL; Sigma-Aldrich, USA), L-glutamine (2 mM; Sigma-Aldrich, USA), hydrocortisone (1 μg/mL; Sigma-Aldrich, USA), heparin (25 U; Sigma-Aldrich, USA), and endothelial growth factor (30 μg/mL; Sigma-Aldrich, USA).

Techniques: Permeability

Journal: Cells

Article Title: The Role of Aldosterone in Vascular Permeability in Diabetes

doi: 10.3390/cells15010089

Figure Lengend Snippet: Mineralocorticoid receptor–dependent mechanisms of diabetic skin vascular permeability and their modulation by eplerenone. ( a ) Experimental diabetes and hyperglycemia are associated with increased aldosterone (ALDO) levels and reduced expression of 11β-hydroxysteroid dehydrogenase type 2 (HSD11β2) in the skin, leading to loss of mineralocorticoid receptor (MR) selectivity and enhanced MR activation by both mineralocorticoids and glucocorticoids. MR overactivation triggers genomic and non-genomic signaling pathways that promote inflammation, oxidative stress, vascular endothelial growth factor (VEGF) up-regulation, and endothelial activation characterized by increased von Willebrand factor (vWF) exocytosis. These processes converge to destabilize endothelial junctions, resulting in endothelial barrier dysfunction and increased skin microvascular permeability, which contributes to the development of diabetic skin disorders. ( b ) Pharmacological MR blockade with eplerenone (EPL) attenuates diabetes-induced skin vascular permeability. EPL reduces MR-dependent inflammatory signaling, oxidative stress, VEGF expression, and vWF exocytosis, thereby improving endothelial barrier function and limiting vascular leakage. These protective effects occur independently of changes in blood pressure or glycemic control, highlighting the pleiotropic vasculoprotective actions of EPL in diabetic skin microvasculature.

Article Snippet: Immortalized human dermal microvascular endothelial cells (HMEC-1; ATCC ® CRL-3243TM; ATCC, Glasgow, UK) were maintained at 37 °C in a 5% CO 2 atmosphere in MCB131 medium (GibcoTM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum, streptomycin–penicillin (100 U/mL; Sigma-Aldrich, USA), L-glutamine (2 mM; Sigma-Aldrich, USA), hydrocortisone (1 μg/mL; Sigma-Aldrich, USA), heparin (25 U; Sigma-Aldrich, USA), and endothelial growth factor (30 μg/mL; Sigma-Aldrich, USA).

Techniques: Permeability, Expressing, Activation Assay, Protein-Protein interactions, Control