Journal: Journal of pediatric gastroenterology and nutrition

Article Title: MATRIX METALLOPROTEINASES IN THE URINE AND TISSUE OF PATIENTS WITH JUVENILE POLYPS: POTENTIAL BIOMARKERS FOR THE PRESENCE OF POLYPS

doi:

Figure Lengend Snippet: Substrate gel electrophoresis (zymography) analysis was performed on the urine from subjects with polyps and healthy age and sex matched controls. Urine was electrophoresed in gelatin-embedded polyacrylamide gels. Gelatin is the substrate for MMP-2 and MMP-9. Neutrophil Gelatinase Associated Lipocalin (NGAL) is a protein that has been found in the urine of polyp patients and migrates in complex with MMP-9 at approximately 125 kDa.

Article Snippet: MMP-2 levels were determined using a Human MMP-2 ELISA kit (R & D Systems, Inc.) and was performed according to the manufacturer’s instructions.

Techniques: Nucleic Acid Electrophoresis, Zymography

Journal: Journal of pediatric gastroenterology and nutrition

Article Title: MATRIX METALLOPROTEINASES IN THE URINE AND TISSUE OF PATIENTS WITH JUVENILE POLYPS: POTENTIAL BIOMARKERS FOR THE PRESENCE OF POLYPS

doi:

Figure Lengend Snippet: Quantitative analysis of MMP-2 by ELISA showed significantly elevated levels of MMP-2 in the urine of polyp subjects as compared with the urine of age-and sex matched controls (medians: 0.046 vs. 0 ng MMP-2/ug total protein, p < 0.001, Mann-Whitney U-test).

Article Snippet: MMP-2 levels were determined using a Human MMP-2 ELISA kit (R & D Systems, Inc.) and was performed according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Journal of pediatric gastroenterology and nutrition

Article Title: MATRIX METALLOPROTEINASES IN THE URINE AND TISSUE OF PATIENTS WITH JUVENILE POLYPS: POTENTIAL BIOMARKERS FOR THE PRESENCE OF POLYPS

doi:

Figure Lengend Snippet: Immunohistochemistry was performed on polyp tissue and normal colonic tissue. Antibodies specific for CD34, MMP-2, MMP-9, and NGAL were used. CD34 staining showed increased microvessel density in polyp tissue. MMP-2 expression was increased in both the lamina propria and the epithelium of polyp tissue compared to normal colonic tissue. MMP-9 expression was higher in the lamina propria and endothelium of polyp tissue. NGAL expression was increased in both the lamina propria and epithelium of polyp tissue compared to normal colonic tissue.

Article Snippet: MMP-2 levels were determined using a Human MMP-2 ELISA kit (R & D Systems, Inc.) and was performed according to the manufacturer’s instructions.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Journal of pediatric gastroenterology and nutrition

Article Title: MATRIX METALLOPROTEINASES IN THE URINE AND TISSUE OF PATIENTS WITH JUVENILE POLYPS: POTENTIAL BIOMARKERS FOR THE PRESENCE OF POLYPS

doi:

Figure Lengend Snippet: Protein Tissue Expression as Determined by polyp Immunohistochemistry

Article Snippet: MMP-2 levels were determined using a Human MMP-2 ELISA kit (R & D Systems, Inc.) and was performed according to the manufacturer’s instructions.

Techniques: Expressing, Immunohistochemistry, Biomarker Assay

Journal: Acta Histochemica et Cytochemica

Article Title: Matrix Metalloproteinase-2 Activated by Ultraviolet-B Degrades Human Ciliary Zonules In Vitro

doi: 10.1267/ahc.20-00021

Figure Lengend Snippet: UV-B irradiation increased matrix metalloproteinase-2 (MMP-2) in cell lysates. Human nonpigmented ciliary epithelial cells were cultured for 4 weeks and then irradiated with UV-B at levels of 0, 50, 100, and 150 mJ/cm 2 . Then, the cells were cultured for another 24 hr under the same culture conditions. Next, cell/matrix layers were collected at 12 ( A ) and 24 ( B ) hr after UV irradiation and analyzed by ELISA. The level of active MMP-2 increased depending on the irradiation level of UV-B. ( A ) The levels of total and active MMP-2 at 150 mJ/cm 2 were significantly increased compared to the control (0 mJ/cm 2 ). ( A ) The levels of total and active MMP-2 at 100 and 150 mJ/cm 2 were significantly increased compared to the control (0 mJ/cm 2 ). ( B ) The level of active MMP-2 at 100 mJ/cm 2 were significantly increased compared to the control (0 mJ/cm 2 ). The amounts of total and active MMP-2 at 150 mJ/cm 2 were significantly increased compared to the control (0 mJ/cm 2 ). The values for each control sample (0 mJ/cm 2 ) were arbitrarily assigned a value of 1. The results represent the mean ± standard deviation of four independent determinations. ** P < 0.05 versus control.

Article Snippet: A human MMP-2 ELISA kit (QuickZyme Biosciences, Leiden, Netherland) was used to measure the protein level of both active-type MMP-2 and total MMP-2 in the cell/matrix of the culture.

Techniques: Irradiation, Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Acta Histochemica et Cytochemica

Article Title: Matrix Metalloproteinase-2 Activated by Ultraviolet-B Degrades Human Ciliary Zonules In Vitro

doi: 10.1267/ahc.20-00021

Figure Lengend Snippet: Inhibitory effect by addition of MMP-2 inhibitor. Human nonpigmented ciliary epithelial cells were cultured for 4 weeks and then irradiated with UV-B at levels of 0, 50, 100, and 150 mJ/cm 2 . Then, 1.5 μM MMP-2 inhibitor was added to the medium for an additional 24 hr. After 24 hr, the cells were simultaneously labeled for fibrillin-1 (green; first panels) and fibrillin-2 (red; second panels). Superimposition of both labels is shown in the third panels. DAPI for nuclear staining is shown in the fourth panels. Bar = 50 μm.

Article Snippet: A human MMP-2 ELISA kit (QuickZyme Biosciences, Leiden, Netherland) was used to measure the protein level of both active-type MMP-2 and total MMP-2 in the cell/matrix of the culture.

Techniques: Cell Culture, Irradiation, Labeling, Staining

Journal: Acta Histochemica et Cytochemica

Article Title: Matrix Metalloproteinase-2 Activated by Ultraviolet-B Degrades Human Ciliary Zonules In Vitro

doi: 10.1267/ahc.20-00021

Figure Lengend Snippet: Inhibitory effect by addition of MMP-2/MMP-9 inhibitor. Human nonpigmented ciliary epithelial cells were cultured for 4 weeks and then irradiated with UV-B at levels of 0, 50, 100, and 150 mJ/cm 2 . Then, 50 nM MMP-2/MMP-9 inhibitor was added to the medium for an additional 24 hr. After 24 hr, the cells were doubly labeled for fibrillin-1 (green; first panels) and fibrillin-2 (red; second panels). Superimposition of both labels is shown in the third panels. DAPI for nuclear staining is shown in the fourth panels. Bar = 50 μm.

Article Snippet: A human MMP-2 ELISA kit (QuickZyme Biosciences, Leiden, Netherland) was used to measure the protein level of both active-type MMP-2 and total MMP-2 in the cell/matrix of the culture.

Techniques: Cell Culture, Irradiation, Labeling, Staining

Journal: Acta Histochemica et Cytochemica

Article Title: Matrix Metalloproteinase-2 Activated by Ultraviolet-B Degrades Human Ciliary Zonules In Vitro

doi: 10.1267/ahc.20-00021

Figure Lengend Snippet: UV-B irradiation increases the level of MMP-2 via p38 activation. Human nonpigmented ciliary epithelial cells were cultured for 24 hr and then the cells pretreated with SB203580 (p38 inhibitor) were irradiated with UV-B at levels of 0, 50, 100, and 150 mJ/cm 2 . Then, the cells were cultured for another 12 and 24 hr under serum-free culture conditions. ( A ) Cell/matrix layers were collected 10 min after UV irradiation and analyzed by Western blot against phosphor-p38. The level of phosphor-p38 treated with SB203580 was inhibited at the same level as the control (0 mJ/cm 2 ). ( B ) Immunofluorescence showed that phosphor-p38 was labeled in the cells at 150 mJ/cm 2 , but not in SB203580-treated cells. Cell/matrix layers were collected at 12 ( C ) and 24 ( D ) hours after UV irradiation and analyzed by ELISA. The active MMP-2 in the SB203580-treated cells was suppressed compared with the SB203580-untreated vehicle cells. The values for each control sample (0 mJ/cm 2 ) without SB203580 were arbitrarily assigned a value of 1. The results represent the mean ± standard deviation of four independent determinations. ** P < 0.05 versus control.

Article Snippet: A human MMP-2 ELISA kit (QuickZyme Biosciences, Leiden, Netherland) was used to measure the protein level of both active-type MMP-2 and total MMP-2 in the cell/matrix of the culture.

Techniques: Irradiation, Activation Assay, Cell Culture, Western Blot, Immunofluorescence, Labeling, Enzyme-linked Immunosorbent Assay, Standard Deviation