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recombinant human hpse  (R&D Systems)


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    R&D Systems recombinant human hpse
    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 <t>HPSE</t> (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.
    Recombinant Human Hpse, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+active+heparanase/Recombinant+Human+Active+Heparanase%2FHPSE+Protein%2C+CF/pmc12974118-120-10-16
    Average 94 stars, based on 29 article reviews
    recombinant human hpse - by Bioz Stars, 2026-10
    94/100 stars

    Images

    1) Product Images from "Elevated vaginal heparan sulfate correlates with impaired neutrophil killing of Candida albicans in women with vulvovaginal candidiasis"

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

    Journal: Infection and Immunity

    doi: 10.1128/iai.00709-25

    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.
    Figure Legend 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.

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

    Related Articles

    Recombinant:

    Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope
    Article Snippet: .. NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570-GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. ..

    Article Title: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice
    Article Snippet: .. The recombinant VEGF 165 , VEGF 121 and biotinylated-VEGF 165 (bt-VEGF 165 ), VEGFR-1, VEGFR-2 and recombinant Human Active Heparanase (HPSE; 7570-GH) were from R&D Systems Inc. (Wiesbaden-Nordenstadt, Germany). .. Heparin sodium salt from porcine intestinal mucosa and Calcium dobesilate (2,5-Dihydroxybenzenesulfonic acid calcium salt), Fluorescein isothiocyanate-dextran, molecular mass: 70 kDa, Duolink ® In Situ PLA kit and probes, calcein-AM and streptozotozin (STZ) were from Sigma Aldrich (Taufkirchen, Germany).

    Article Title: Netrin-1 inhibits the attachment and internalization of Hepatitis B virus for hepatocyte infection
    Article Snippet: .. Recombinant Human Active Heparanase (7570-GH-005; R&D Systems, Minneapolis, MN) at a final concentration of 0.5 μg/mL was added to the cells followed by incubation at 37°C for 1.5 h. After incubation, the medium was removed and the cells were used in the next step of the HBV attachment assay. .. HBV DNA was extracted from the cells using a DNeasy Blood & Tissue Kit (QIAGEN, Valencia, CA).

    Article Title: Loss of VEGFB and its signaling in the diabetic heart is associated with increased cell death signaling.
    Article Snippet: Anti-vinculin (13901), anti-phospho ERK 175 (4370), anti-total ERK (9102), anti-phospho GSK3β (9336), which detects serine 9 176 phosphorylation, anti-GSK3β (9315), anti-PARP (9542) and anti-caspase 3 (9662) 177 antibodies were obtained from Cell Signaling Technology. .. Purified Hep-L was prepared 178 as described previously (47), recombinant human active heparanase (7570-GH-005) 179 was bought from R&D Systems, and heparin (Hepalean, 1000U/ml) was purchased 180 from Organon, Canada. .. For Western blots that detect only Hep-L, we used the 181 heparanase (N-Term) antibody (ABIN786265), which preferentially recognizes the 65 182 kDa Hep-L, from Aviva Systems Biology.

    Article Title: Netrin-1 inhibits the attachment and internalization of Hepatitis B virus for hepatocyte infection.
    Article Snippet: .. Recombinant Human Active Heparanase (7570-GH-005; R&D Systems, Minneapolis, MN) at a final concentration of 0.5 μg/mL was added to the cells followed by incubation at 37°C for 1.5 h. After incubation, the medium was removed and the cells were used in the next step of the HBV attachment assay. .. HBV DNA was extracted from the cells using a DNeasy Blood & Tissue Kit (QIAGEN, Valencia, CA).

    Article Title: Heparanase inhibitors and use thereof
    Article Snippet: .. Recombinant human active heparanase derived from Chinese hamster ovary cells was from R&D Systems. .. Bovine serum albumin-coated 96 well microplates (96F Maxisorp NNC #456537, Thermo Scientific) were used for the assays and were prepared by incubation of the plates with 1% (w/v) BSA dissolved in phosphate-buffered saline containing 0.05% (v/v) Tween-20 (PBST) at 37° C. for 1 h. The plates were washed three times with PBST, shaken dry, and stored for up to two weeks at 4° C. before use.

    Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope
    Article Snippet: Heparanase treatment of neuroblastoma cell lines NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570- GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. .. Heparanase treatment of neuroblastoma cell lines NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570- GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. .. RNA sequencing data from SCLC tumors and cell lines and paired normal lung tissues was accessed via the GEO database accession number GSE60052 28 and European Genome-phenome Archive from the accession number EGAS00001000334 from data generated by Genentech Research and Early Development29 and both reprocessed at CHOP as described below.

    Article Title: Heparanase Activates Antithrombin through the Binding to Its Heparin Binding Site.
    Article Snippet: Culture media were obtained from Gibco (Fisher Scientific, Madrid, Spain). .. Recombinant human active heparanase was from R&D systems. .. TFPI presence on heparanase preparation was evaluated by silver staining and immunostaining with anti-human TFPI (from rabbit, Santa Cruz Biotechnologies, USA) followed by donkey anti-rabbit IgG–horseradish peroxidase conjugate (GE Healthcare, Barcelona, Spain), with detection via an ECL kit (GE Healthcare, Barcelona, Spain).

    Western Blot:

    Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope
    Article Snippet: .. NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570-GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. ..

    Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope
    Article Snippet: Heparanase treatment of neuroblastoma cell lines NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570- GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. .. Heparanase treatment of neuroblastoma cell lines NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570- GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. .. RNA sequencing data from SCLC tumors and cell lines and paired normal lung tissues was accessed via the GEO database accession number GSE60052 28 and European Genome-phenome Archive from the accession number EGAS00001000334 from data generated by Genentech Research and Early Development29 and both reprocessed at CHOP as described below.

    Staining:

    Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope
    Article Snippet: .. NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570-GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. ..

    Article Title: A GPC2 antibody-drug conjugate is efficacious against neuroblastoma and small-cell lung cancer via binding a conformational epitope
    Article Snippet: Heparanase treatment of neuroblastoma cell lines NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570- GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. .. Heparanase treatment of neuroblastoma cell lines NB-EbC1, NB-SD, and NB-1643 cell lines were treated with 150 ng/mL recombinant human active heparanase (R&D Systems, 7570- GH-005) for 2 or 18 hours at which point cells were analyzed by western blot and stained with the D3-GPC2-IgG1-PE antibody as above. .. RNA sequencing data from SCLC tumors and cell lines and paired normal lung tissues was accessed via the GEO database accession number GSE60052 28 and European Genome-phenome Archive from the accession number EGAS00001000334 from data generated by Genentech Research and Early Development29 and both reprocessed at CHOP as described below.

    Concentration Assay:

    Article Title: Netrin-1 inhibits the attachment and internalization of Hepatitis B virus for hepatocyte infection
    Article Snippet: .. Recombinant Human Active Heparanase (7570-GH-005; R&D Systems, Minneapolis, MN) at a final concentration of 0.5 μg/mL was added to the cells followed by incubation at 37°C for 1.5 h. After incubation, the medium was removed and the cells were used in the next step of the HBV attachment assay. .. HBV DNA was extracted from the cells using a DNeasy Blood & Tissue Kit (QIAGEN, Valencia, CA).

    Article Title: Netrin-1 inhibits the attachment and internalization of Hepatitis B virus for hepatocyte infection.
    Article Snippet: .. Recombinant Human Active Heparanase (7570-GH-005; R&D Systems, Minneapolis, MN) at a final concentration of 0.5 μg/mL was added to the cells followed by incubation at 37°C for 1.5 h. After incubation, the medium was removed and the cells were used in the next step of the HBV attachment assay. .. HBV DNA was extracted from the cells using a DNeasy Blood & Tissue Kit (QIAGEN, Valencia, CA).

    Incubation:

    Article Title: Netrin-1 inhibits the attachment and internalization of Hepatitis B virus for hepatocyte infection
    Article Snippet: .. Recombinant Human Active Heparanase (7570-GH-005; R&D Systems, Minneapolis, MN) at a final concentration of 0.5 μg/mL was added to the cells followed by incubation at 37°C for 1.5 h. After incubation, the medium was removed and the cells were used in the next step of the HBV attachment assay. .. HBV DNA was extracted from the cells using a DNeasy Blood & Tissue Kit (QIAGEN, Valencia, CA).

    Article Title: Netrin-1 inhibits the attachment and internalization of Hepatitis B virus for hepatocyte infection.
    Article Snippet: .. Recombinant Human Active Heparanase (7570-GH-005; R&D Systems, Minneapolis, MN) at a final concentration of 0.5 μg/mL was added to the cells followed by incubation at 37°C for 1.5 h. After incubation, the medium was removed and the cells were used in the next step of the HBV attachment assay. .. HBV DNA was extracted from the cells using a DNeasy Blood & Tissue Kit (QIAGEN, Valencia, CA).

    Purification:

    Article Title: Loss of VEGFB and its signaling in the diabetic heart is associated with increased cell death signaling.
    Article Snippet: Anti-vinculin (13901), anti-phospho ERK 175 (4370), anti-total ERK (9102), anti-phospho GSK3β (9336), which detects serine 9 176 phosphorylation, anti-GSK3β (9315), anti-PARP (9542) and anti-caspase 3 (9662) 177 antibodies were obtained from Cell Signaling Technology. .. Purified Hep-L was prepared 178 as described previously (47), recombinant human active heparanase (7570-GH-005) 179 was bought from R&D Systems, and heparin (Hepalean, 1000U/ml) was purchased 180 from Organon, Canada. .. For Western blots that detect only Hep-L, we used the 181 heparanase (N-Term) antibody (ABIN786265), which preferentially recognizes the 65 182 kDa Hep-L, from Aviva Systems Biology.

    Derivative Assay:

    Article Title: Heparanase inhibitors and use thereof
    Article Snippet: .. Recombinant human active heparanase derived from Chinese hamster ovary cells was from R&D Systems. .. Bovine serum albumin-coated 96 well microplates (96F Maxisorp NNC #456537, Thermo Scientific) were used for the assays and were prepared by incubation of the plates with 1% (w/v) BSA dissolved in phosphate-buffered saline containing 0.05% (v/v) Tween-20 (PBST) at 37° C. for 1 h. The plates were washed three times with PBST, shaken dry, and stored for up to two weeks at 4° C. before use.



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    94
    R&D Systems cf catalog 7570 gh 005 r d systems
    In vitro glycocalyx degradation mediated by <t>heparanase</t> 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, <t>recombinant</t> human syndecan-1; MMP-9, matrix metalloproteinase 9; HPSE, human heparanase.
    Cf Catalog 7570 Gh 005 R D Systems, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+active+heparanase/Recombinant+Human+Active+Heparanase%2FHPSE+Protein%2C+CF/pmc12276975-978-5-9
    Average 94 stars, based on 1 article reviews
    cf catalog 7570 gh 005 r d systems - by Bioz Stars, 2026-10
    94/100 stars
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    recombinant human active heparanase hpse protein  (R&D Systems)
    94
    R&D Systems recombinant human active heparanase hpse protein
    In vitro glycocalyx degradation mediated by <t>heparanase</t> 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, <t>recombinant</t> human syndecan-1; MMP-9, matrix metalloproteinase 9; HPSE, human heparanase.
    Recombinant Human Active Heparanase Hpse Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/recombinant+human+active+heparanase/Recombinant+Human+Active+Heparanase%2FHPSE+Protein%2C+CF/pmc12276975-978-0-9
    Average 94 stars, based on 1 article reviews
    recombinant human active heparanase hpse protein - by Bioz Stars, 2026-10
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    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.

    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

    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.

    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

    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.

    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