Journal: Translational Vision Science & Technology

Article Title: Differentiating Ocular Pathogenic Staphylococcus epidermidis Isolates Using a Human Corneal Epithelial Cell Coculture Model

doi: 10.1167/tvst.14.11.21

Figure Lengend Snippet: Establishment and optimization of a coculture model using HCECs and S. epidermidis . The model was developed using three representative S. epidermidis strains: a reference strain (ATCC 35984), a commensal isolate from a normal conjunctiva, and a clinical isolate from a patient with keratitis. ( A ) Time-course analysis showing phase-contrast images of HCECs cocultured with the three strains at 10⁷ CFU/mL for up to 72 hours. ( B ) Dose–response analysis showing phase-contrast images of HCECs after 24 hours of coculture with various concentrations (10⁴ to 10⁸ CFU/mL) of the strains. ( C ) Corresponding fluorescence microscopy images of HCECs stained with calcein AM (green fluorescence for live cells) to assess cell viability under the same dose–response conditions. ( D–F ) Quantification of HCEC viability from the calcein AM assay following 24-hour treatment with ( D ) ATCC 35984, ( E ) normal conjunctiva, and ( F ) keratitis strains. Scale bars , 100 µm. * P < 0.01; ** P < 0.01; *** P < 0.001; **** P < 0.0001; calcein AM, calcein acetoxymethyl ester; NC, normal conjunctiva; ns, no significance.

Article Snippet: ATCC 35984 induced apparent HCEC destruction at concentrations of 10 6 , 10 7 , and 10 8 CFU/mL in the phase-contrast set ( B).

Techniques: Fluorescence, Microscopy, Staining, Calcein AM Assay

Journal: Translational Vision Science & Technology

Article Title: Differentiating Ocular Pathogenic Staphylococcus epidermidis Isolates Using a Human Corneal Epithelial Cell Coculture Model

doi: 10.1167/tvst.14.11.21

Figure Lengend Snippet: HCEC viability after exposure to 17 S. epidermidis strains at varying concentrations. ( A ) Nonpathogenic strains. ( B, C ) Pathogenic strains. Asterisks indicate a significant decrease in cell viability compared with the PBS control. * P < 0.05; ** P < 0.01; *** P < 0.001; *** P < 0.0001.

Article Snippet: ATCC 35984 induced apparent HCEC destruction at concentrations of 10 6 , 10 7 , and 10 8 CFU/mL in the phase-contrast set ( B).

Techniques: Control

Journal: Translational Vision Science & Technology

Article Title: Differentiating Ocular Pathogenic Staphylococcus epidermidis Isolates Using a Human Corneal Epithelial Cell Coculture Model

doi: 10.1167/tvst.14.11.21

Figure Lengend Snippet: HCEC viability distinguished pathogenic from nonpathogenic S. epidermidis strains. ( A ) Heatmap illustrating HCEC viability following exposure to varying concentrations of 12 pathogenic and 5 nonpathogenic strains. Comparison of HCEC viability after exposure to ( B ) 104, ( C ) 105, ( D ) 106, ( E ) 107, and ( F ) 108 CFU/mL of pathogenic and nonpathogenic groups. ROC analysis of the diagnostic performance of HCEC viability at ( G ) 104, ( H ) 105, ( I ) 106, ( J ) 107, and ( K ) 108 CFU/mL in distinguishing the pathogenic S. epidermidis strains. ns, not significant. ** P < 0.01.

Article Snippet: ATCC 35984 induced apparent HCEC destruction at concentrations of 10 6 , 10 7 , and 10 8 CFU/mL in the phase-contrast set ( B).

Techniques: Comparison, Diagnostic Assay

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F expression is regulated by hypoxia in cardiac endothelial cells. (A) The expression of Sema3F RNA was increased in endothelial cells (HCECs) under hypoxic conditions, with the highest levels being observed after 4 h. HCECs were exposed to hypoxia (1 % O 2 ) for the indicated times, after which the cells were harvested, RNA was isolated and reverse transcribed. cDNA was then used for the quantification of Sema3F expression by qRT-PCR. The data presented here is derived from four independent experiments (biological replicates, n = 4), and the statistical analysis employed was the Man-Whitney test. A p-value <0.05 is considered to be significant and marked with an asterisk (*). (B) Sema3F protein was significantly increased in response to 24 h of hypoxia in comparison to normoxia. For Western blot analysis HCECs were lysed and Sema3F protein levels were detected by an anti-Sema3F antibody. GAPDH was used as a loading control. Representative blots from four independent experiments are presented (biological replicates, n = 4). The results were analyzed using the Mann-Whitney test (*p < 0.05).

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Expressing, Isolation, Reverse Transcription, Quantitative RT-PCR, Derivative Assay, Comparison, Western Blot, Control, MANN-WHITNEY

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F is an inhibitor of angiogenesis in Matrigel® sprouting assays. (A-C) In cardiac endothelial cells (HCECs) Sema3F abolished the pro-angiogenic effect of VEGF. HCECs were cultured with the indicated amounts of Sema3F or VEGF alone or in combination over 16 to 18 h. HCECs were then assayed on Matrigel® for further 2.5 h. (A) Representative micrographs of stimulated HCECs are shown. (B&C) Sema3F prevented VEGF-induced cumulative sprout length (B) and branching of HCECs (C). Data from 4 independent experiments were shown (n, number of biological replicates). The results were analyzed using the one-way ANOVA and Bonferroni's multiple comparisons test: * p < 0.05. Figs. D—F show the effect of Sema3F knockdown on angiogenesis of HCECs. The siRNA-based Sema3F knockdown resulted in a potentiated pro-angiogenic effect of VEGF. HCECs transfected with Sema3F-specific siRNA (siSema3F) or control siRNA (ctrl-siRNA) were treated with 100 ng/ml VEGF or EBM with 2 % FCS for 18 h before being assayed on Matrigel®. (D) Representative micrographs are shown. Sema3F knockdown led to a significant increase in cumulative sprout length (E) and number of branch points (F). (E&F) Data from 4 independent experiments were shown (n, number of biological replicates). All box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using one-way ANOVA followed by Bonferroni's multiple comparison test: * p < 0.05.

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Cell Culture, Knockdown, Transfection, Control, Comparison

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Semaphorin 3F is elevated in serum of heart failure patients and inhibits cardiac angiogenesis via the VEGF/Akt/eNOS pathway

doi: 10.1016/j.jmccpl.2025.100470

Figure Lengend Snippet: Sema3F inhibits angiogenesis in spheroid sprouting and aortic ring assay. (A&B) For the Spheroid sprouting assay, HCECs were formed to spheroids and were then stimulated for 24 h with the indicated amounts of Sema3F, VEGF or both. (A) Representative micrographs are shown. (B) Sema3F caused a significant decrease in sprout length and reduced the proangiogenic effect of VEGF. Data from 4 independent experiments are shown. Ten spheroids per well per condition were measured and the average cumulative sprout length was estimated (n, number of biological replicates). Analysis with one-way ANOVA followed by Bonferroni's multiple comparisons test. *p < 0.05. (C&D) In the aortic ring assay, aortic rings were stimulated with the indicated amounts of Sema3F, VEGF or both on days 3, 5 and 7. The number of capillaries was counted on day 9. (C) Compared to controls, VEGF alone increased sprouting, while co-treatment of aortic rings with Sema3F prevented VEGF-induced sprouting ex vivo. Data from 3 independent experiments, total of 10–12 aortic rings per group (n, number of biological replicates). All box plots extend from the 25th to the 75th percentile with the median marked. Whiskers extend from the 10th to the 90th percentile. The results were analyzed using one-way ANOVA followed by Bonferroni's multiple comparisons test. * p < 0.05.

Article Snippet: Human cardiac endothelial cells (HCECs) were purchased from PELOBIOTECH GmbH, Planegg, DE and were grown at 37 °C, 5 % CO 2 in growth factor enhanced microvascular endothelial cell medium (PELOBIOTECH GmbH, Planegg, DE) supplemented with 10 % fetal calf serum (FCS) obtained from Lonza Pharma & Biotech (Verviers, BEL).

Techniques: Aortic Ring Assay, Ex Vivo