Journal: Infection and Immunity

Article Title: Elevated vaginal heparan sulfate correlates with impaired neutrophil killing of Candida albicans in women with vulvovaginal candidiasis

doi: 10.1128/iai.00709-25

Figure Lengend Snippet: PMN antifungal activity in VCM spiked with HS or treated with HPSEs. Vaginal swab samples were collected from three groups: women with RVVC experiencing a symptomatic acute episode of VVC (symptomatic group, n = 31), those in asymptomatic remission (asymptomatic group, n = 14), and healthy women without a history of frequent VVC (control group, n = 16). The data sets included an additional five follow-up swabs collected from women in the symptomatic and asymptomatic groups. Detached swab tips were suspended individually in 1 mL RPMI 1640 medium to release vaginal secretions and cellular content. Supernatants were sterile-filtered as VCM. PMNs (5 × 10 5 ) isolated from peripheral blood of healthy volunteers were incubated with C. albicans cells (1 × 10 5 ) in 100 µL of VCM or RPMI medium for 3 h at 37°C with 5% CO 2 . Controls consisted of C. albicans cultured alone in VCM or RPMI medium and were used to calculate % killing. ( A ) VCM samples with normal killing activity (non-inhibitory VCM, gray bar) were spiked with purified porcine HS (500 µg/mL) and reevaluated for PMN killing activity (blue bar). Spiked non-inhibitory VCM was then pretreated with r Ph HPSE (3 ng/mL) and reevaluated for PMN killing activity (red bar). ( B ) VCM samples with low killing activity (inhibitory VCM, solid blue bar) were pretreated with rhHPSE (4 µg/mL, red-dotted bar) or r Ph HPSE (3 ng/mL, red-striped bar) and reevaluated for PMN killing activity. After coculture, viable C. albicans cells were enumerated by quantitative plate counts. Data were analyzed using the one-way ANOVA and Bonferroni’s post-test. Bar heights and error bars represent group means ± SEM for % killing values across independent replicates of unique VCM samples. The graphs represent cumulative data from three to four experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: To degrade HS, symptomatic (inhibitory) VCM was pretreated with either recombinant human HPSE (rhHPSE, 4 μg/mL, R&D Systems) or recombinant Pedobacter heparinus heparinase III (r Ph HPSE, 3 ng/mL, R&D Systems) for 1 h at 37°C.

Techniques: Activity Assay, Control, Sterility, Isolation, Incubation, Cell Culture, Purification

Journal: iScience

Article Title: Matrix metalloproteinase-9 mediates endothelial glycocalyx degradation and correlates with severity of hemorrhagic fever with renal syndrome

doi: 10.1016/j.isci.2025.113262

Figure Lengend Snippet: In vitro glycocalyx degradation mediated by heparanase and MMP-9 followed by QCM-D (A) b-HS and (B) rhSDC-1 protein were absorbed on synthetic membrane composed by POPC-DOPE (95:5) lipids and streptavidin, plus Tris-NTA linker for rhSDC-1. After incubation with albumin at 40, 20, or 10 mg/mL, real-time degradation of b-HS by HPSE at 250 ng/mL (C) and rhSDC1 by MMP-9 at 500 ng/mL (D). Arrow indicates when the enzyme is injected. Shift in frequency (Δf) and dissipation (ΔD) due to enzyme activity was measured after enzyme activity in enzyme buffer. Percentage of Δf and ΔD signal reduction for b-HS (E) and rhSDC1 (F) after enzymatic treatment ( n = 4) were calculated from the b-HS or rhSDC-1 signal in enzyme buffer (corresponding to 100%). Data are represented as mean ± SD. Significant differences are indicated by asterisks (∗∗∗ p < 0.001; ∗∗ p < 0.01; and ∗ p < 0.05). QCM-D, quartz crystal microbalance with dissipation; b-HS, biotinylated heparan sulfate; rhSDC-1, recombinant human syndecan-1; MMP-9, matrix metalloproteinase 9; HPSE, human heparanase.

Article Snippet: Recombinant human heparanase (rhHPSE; #7570-GH-005, R&D Systems, Sweden) was diluted in rhHPSE assay buffer comprising 20 mM Tris-HCl and 4 mM CaCl 2 (pH 7.5).

Techniques: In Vitro, QCM-D, Membrane, Incubation, Injection, Activity Assay, Recombinant

Journal: iScience

Article Title: Matrix metalloproteinase-9 mediates endothelial glycocalyx degradation and correlates with severity of hemorrhagic fever with renal syndrome

doi: 10.1016/j.isci.2025.113262

Figure Lengend Snippet: In vitro endothelial glycocalyx degradation and endothelial barrier disruption followed by impedance measurement (A) SDC-1 staining on ciGEnC cells after incubation with MMP-9 enzyme and increased concentration of albumin (scale bars 20 μm). (B) SDC-1 fluorescence quantification after treatment compared to control (no treatment) (averaged values of 3 sections per condition, n = 3 assays). (C) HS staining on ciGEnC cells after incubation with HSPE enzyme and increased concentration of albumin (scale bars 20 μm). (D) HS fluorescence quantification after treatment compared to control (no treatment) (averaged values of 3 sections per condition, n = 3 assays). (E) Endothelial barrier resistance followed by impedance measurement (ECIS, APBiophysic) on ciGEnC after treatment with MMP-9 and albumin (averaged of 2 values per conditions, n = 3 assays). (F) Endothelial barrier resistance followed by impedance measurement (ECIS, APBiophysic) on ciGEnC after treatment with HSPE and albumin (averaged of 2 values per conditions, n = 3 assays). Data are represented as mean ± SD. Significant differences are indicated by asterisks (∗∗∗ p < 0.001; ∗∗ p < 0.01; and ∗ p < 0.05). HS, heparan sulfate; SDC-1, syndecan-1; MMP-9, matrix metalloproteinase 9; HPSE, human heparanase.

Article Snippet: Recombinant human heparanase (rhHPSE; #7570-GH-005, R&D Systems, Sweden) was diluted in rhHPSE assay buffer comprising 20 mM Tris-HCl and 4 mM CaCl 2 (pH 7.5).

Techniques: In Vitro, Disruption, Staining, Incubation, Concentration Assay, Fluorescence, Control