Journal: The Journal of International Medical Research

Article Title: Osteonectin as a screening marker for pancreatic cancer: A prospective study

doi: 10.1177/0300060518772413

Figure Lengend Snippet: Osteonectin serum concentrations in ng/mL in patients with cancer, patients with acute pancreatitis, and healthy individuals

Article Snippet: Human osteonectin was detected by a quantitative sandwich enzyme immunoassay technique (Human Osteonectin ELISA kit; Cusabio, College Park, MD, USA) according to the manufacturer’s instructions.

Techniques:

Journal: The Journal of International Medical Research

Article Title: Osteonectin as a screening marker for pancreatic cancer: A prospective study

doi: 10.1177/0300060518772413

Figure Lengend Snippet: Box plot by group shows the median, 25th to 75th interquartile range, and minimum to maximum serum osteonectin concentrations in ng/mL.

Article Snippet: Human osteonectin was detected by a quantitative sandwich enzyme immunoassay technique (Human Osteonectin ELISA kit; Cusabio, College Park, MD, USA) according to the manufacturer’s instructions.

Techniques:

Journal: The Journal of International Medical Research

Article Title: Osteonectin as a screening marker for pancreatic cancer: A prospective study

doi: 10.1177/0300060518772413

Figure Lengend Snippet: Receiver operating characteristic curve. Serum osteonectin concentrations of >100.18 ng/mL at enrolment had 86% accuracy in predicting pancreatic cancer.

Article Snippet: Human osteonectin was detected by a quantitative sandwich enzyme immunoassay technique (Human Osteonectin ELISA kit; Cusabio, College Park, MD, USA) according to the manufacturer’s instructions.

Techniques:

Journal: The Journal of International Medical Research

Article Title: Osteonectin as a screening marker for pancreatic cancer: A prospective study

doi: 10.1177/0300060518772413

Figure Lengend Snippet: Least-square means and 95% confidence intervals of serum osteonectin concentrations in ng/mL among the three groups

Article Snippet: Human osteonectin was detected by a quantitative sandwich enzyme immunoassay technique (Human Osteonectin ELISA kit; Cusabio, College Park, MD, USA) according to the manufacturer’s instructions.

Techniques:

Journal: Cells

Article Title: In Vivo Effects of Bay 11-7082 on Fibroid Growth and Gene Expression: A Preclinical Study.

doi: 10.3390/cells13131091

Figure Lengend Snippet: Figure 3. Relative mRNA expression levels of SPARC, TDO2, MYD88, TLR6, TLR3, IL6, IL8, TNFα, TNFRSF11A, IL1β, COL3A1, FN1, LOX, TGFB3, CCND1, E2F1, CKS2, PRL, ESR1, PGR, VEGFC, PDGFA and HTR1B in subcutaneously implanted xenografts in ovariectomized CB-17 SCID/Beige mice (n = 10) following 8 weeks of treatment with vehicle or Bay 11-7082 (20 mg/kg/daily). Data are presented as mean ± SEM, with p values indicated on respective lines. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Primary antibodies targeting COL3A1, FN1, SPARC, TDO2, and cleaved caspase-3 were purchased from Proteintech Group, Inc. (Chicago, IL, USA).

Techniques: Expressing

Journal: Cells

Article Title: In Vivo Effects of Bay 11-7082 on Fibroid Growth and Gene Expression: A Preclinical Study.

doi: 10.3390/cells13131091

Figure Lengend Snippet: Figure 4. (A) Representative Western blot analysis of COL3A1, FN1, SPARC, TDO2, and cleaved caspase 3, with corresponding bar graphs (B) showing their relative band densities in the xenografts (n = 10). (C) Total collagen levels assessed by enzyme-linked immunosorbent assay in the 10 xenografts. Data are presented as mean ± SEM of independent experiments, with p values indicated on the respective line. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies targeting COL3A1, FN1, SPARC, TDO2, and cleaved caspase-3 were purchased from Proteintech Group, Inc. (Chicago, IL, USA).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: In Vivo Effects of Bay 11-7082 on Fibroid Growth and Gene Expression: A Preclinical Study.

doi: 10.3390/cells13131091

Figure Lengend Snippet: Figure 6. Relative expression of SPARC, TDO2, MYD88, TLR6, TLR3, IL6, IL8, TNFα, TNFRSF11A, IL1β, COL3A1, FN1, LOX, TGFB3, CCND1, E2F1, CKS2, PRL, ESR1, PGR, VEGFC, PDGFA and HTR1B mRNA in fibroid explants (n = 4) following 48 h of treatment with vehicle or Bay 11-7082 (5 µM). The results are presented as mean ± SEM with p values indicated by corresponding lines. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies targeting COL3A1, FN1, SPARC, TDO2, and cleaved caspase-3 were purchased from Proteintech Group, Inc. (Chicago, IL, USA).

Techniques: Expressing

Journal: Hepatology communications

Article Title: Suboptimal Level of Bone-Forming Cells in Advanced Cirrhosis are Associated with Hepatic Osteodystrophy.

doi: 10.1002/hep4.1234

Figure Lengend Snippet: FIG. 2. Images of BM biopsy in Cirrhosis and Control cases. (A) Panel I. Representative Images of cirrhosis and control BM (left) showing the osteoblast cells and bar graph (right) showing number of osteoblast (mean ± SE) in cirrhosis and control BM (H&E). Panel II. Cirrhosis and control BM (left) showing the osteocytes and bar graph (right) showing number of osteocytes (mean ± SE) in cirrhosis and control BM (H&E). Panel III. Alcian blue-stained chondroblasts in cirrhosis and control BM biopsy section (left) and bar graph (right) showing number of chondroblasts (mean ± SE) in cirrhosis and control BM. (B) Panel I. Representing osteocalcin+ cells (mean ± SE) in cirrhosis and control BM biopsy section (left) and bar graph (right) showing number of osteocalcin+ cells in cirrhosis and control BM. Panel II. Osteocalcin+ area (mean ± SE) in cirrhosis and control BM biopsy section (left) and bar graph (right) showing percentage osteocalcin+ area in cirrhosis and control BM. Panel III. Showing osteonectin+ cells (mean ± SE) in cirrhosis and control BM biopsy section (left) and bar graph (right) showing osteonectin in cirrhosis and control BM (C) Nestin+ BM MSCs in cirrhosis and control BM biopsy section (left) and bar graph (right) showing number of these cells in cirrhosis and control BM. (D) CD169+ BM macrophage (mean ± SE) in cirrhosis and control BM biopsy section (left) and bar graph (right) showing number of CD169+ cells in cirrhosis and control BM. (All image magnification ×200, area 1.3 mm2). Abbreviations: BM, bone marrow; H&E, hematoxylin and eosin.

Article Snippet: Osteocalcin- and osterix+ cell counting were done using Image J software (summarized in the Supporting Material). enzyme-linked immunosorbent assay Enzyme-linked immunosorbent assay for osteocalcin (ABIN368354; N-MID, Aachen, Germany), osteonectin (E-EL-H1351; Elabscience), bone morphogenetic protein 2 (BMP2) (E13650313; Sincere Biotech, Beijing, China), fibroblast growth factor 23 (FGF23) (E13652282; Sincere Biotech), and dipeptidyl peptidase 4 (DPP4) (E-EL-H0058; Elabscience) was performed on BM plasma and peripheral blood plasma of cirrhosis and control cases. skeletal survey A baseline bone survey for bone loss was done by a DEXA scan of the lumbar spine and femoral neck in cases of cirrhosis and was classified as described. (6) statistical analysis Statistical analysis was carried out using Statistical Package for Social Sciences (SPSS, version 22.0; IBM Corp, Armonk, NY).

Techniques: Control, Staining

Journal: Hepatology communications

Article Title: Suboptimal Level of Bone-Forming Cells in Advanced Cirrhosis are Associated with Hepatic Osteodystrophy.

doi: 10.1002/hep4.1234

Figure Lengend Snippet: FIG. 4. Graphs showing BM plasma (left) and peripheral blood plasma (right) of the same patients displaying levels of bone-building proteins. (A) Osteocalcin, (B) osteonectin, (C) bone morphogenic protein, and (D) anti-bone building proteins FGF23 and (E) DPP4 in control and patients with cirrhosis with Child-Turcotte-Pugh (CTP) A, B, or C { median (interquartile range) }.

Article Snippet: Osteocalcin- and osterix+ cell counting were done using Image J software (summarized in the Supporting Material). enzyme-linked immunosorbent assay Enzyme-linked immunosorbent assay for osteocalcin (ABIN368354; N-MID, Aachen, Germany), osteonectin (E-EL-H1351; Elabscience), bone morphogenetic protein 2 (BMP2) (E13650313; Sincere Biotech, Beijing, China), fibroblast growth factor 23 (FGF23) (E13652282; Sincere Biotech), and dipeptidyl peptidase 4 (DPP4) (E-EL-H0058; Elabscience) was performed on BM plasma and peripheral blood plasma of cirrhosis and control cases. skeletal survey A baseline bone survey for bone loss was done by a DEXA scan of the lumbar spine and femoral neck in cases of cirrhosis and was classified as described. (6) statistical analysis Statistical analysis was carried out using Statistical Package for Social Sciences (SPSS, version 22.0; IBM Corp, Armonk, NY).

Techniques: Clinical Proteomics, Control

Journal: Hepatology communications

Article Title: Suboptimal Level of Bone-Forming Cells in Advanced Cirrhosis are Associated with Hepatic Osteodystrophy.

doi: 10.1002/hep4.1234

Figure Lengend Snippet: FIG. 5. Distribution graphs. (A) Panel I. Osteoblasts. Panel II. Osteocytes. Panel III. Chondroblasts. (B) Panel I. Osteocalcin+ cells. Panel II. Percentage osteocalcin+ area. Panel III. Osteonectin+ cells. (C) Nestin+ MSCs. (D) CD169+ BM macrophages (all expressed in mean ± SE) in BM sections of patients with cirrhosis with normal bone density, osteopenia, and osteoporosis; evaluated by T score on bone densitometry scan.

Article Snippet: Osteocalcin- and osterix+ cell counting were done using Image J software (summarized in the Supporting Material). enzyme-linked immunosorbent assay Enzyme-linked immunosorbent assay for osteocalcin (ABIN368354; N-MID, Aachen, Germany), osteonectin (E-EL-H1351; Elabscience), bone morphogenetic protein 2 (BMP2) (E13650313; Sincere Biotech, Beijing, China), fibroblast growth factor 23 (FGF23) (E13652282; Sincere Biotech), and dipeptidyl peptidase 4 (DPP4) (E-EL-H0058; Elabscience) was performed on BM plasma and peripheral blood plasma of cirrhosis and control cases. skeletal survey A baseline bone survey for bone loss was done by a DEXA scan of the lumbar spine and femoral neck in cases of cirrhosis and was classified as described. (6) statistical analysis Statistical analysis was carried out using Statistical Package for Social Sciences (SPSS, version 22.0; IBM Corp, Armonk, NY).

Techniques:

Journal: Cancer Research

Article Title: Macrophage-Derived SPARC Bridges Tumor Cell-Extracellular Matrix Interactions toward Metastasis

doi: 10.1158/0008-5472.can-08-1327

Figure Lengend Snippet: Figure 1. SPARC is required for spontaneous metastasis. 4T1 cells were injected into the mammary fat pad (A and B) or i.v. (C) of wt (.) or SPARC KO (o) mice. A, growth of primary tumor following mammary fat pad injection. Points, mean volume (n = 20); bars, SD. B, spontaneous lung metastases from tumors shown in A. Bars, median number of lung colonies. Clonogenic assay was done 28 d after mammary fat pad cell injection. ***, P < 0.001, two-tailed Mann-Whitney t test. C, experimental lung colonies after i.v. cell injection. Bars, median number of lung metastasis. Clonogenic assay was done 14 d after i.v. cell injection.

Article Snippet: The underside of the Transwell filter inserts (6.5 mm; Costar) were coated with 10 Ag/mL of fibronectin (Sigma), multimeric vitronectin from bovine plasma (Oxford Biomedical Research, UK), rat tail collagen type I (Becton Dickinson), and/or 200 ng of mouse SPARC purified from parietal yolk sac (Sigma) or recombinant human SPARC (ProSci) overnight at 4jC.

Techniques: Injection, Clonogenic Assay, Two Tailed Test, MANN-WHITNEY

Journal: Cancer Research

Article Title: Macrophage-Derived SPARC Bridges Tumor Cell-Extracellular Matrix Interactions toward Metastasis

doi: 10.1158/0008-5472.can-08-1327

Figure Lengend Snippet: Figure 2. Bone marrow–derived SPARC affects 4T1 metastasis in vivo. A, SPARC is barely expressed by neoplastic cells (faint staining of the tumor cell cytoplasm) in 4T1 tumors grown in either wt (a) or SPARC KO (b) mice; additional SPARC expression in wt mice was found in cells that appear as tumor-infiltrating inflammatory cells mostly endowed with widespread cytoplasmic processes that encircled the tumor cells (a and b, 630; inset, 1,000). Accordingly, immunofluorescence and confocal microscopy show that, in tumors from wt mice, SPARC was mostly produced by Mac-1+

Article Snippet: The underside of the Transwell filter inserts (6.5 mm; Costar) were coated with 10 Ag/mL of fibronectin (Sigma), multimeric vitronectin from bovine plasma (Oxford Biomedical Research, UK), rat tail collagen type I (Becton Dickinson), and/or 200 ng of mouse SPARC purified from parietal yolk sac (Sigma) or recombinant human SPARC (ProSci) overnight at 4jC.

Techniques: Derivative Assay, In Vivo, Staining, Expressing, Immunofluorescence, Confocal Microscopy, Produced

Journal: Cancer Research

Article Title: Macrophage-Derived SPARC Bridges Tumor Cell-Extracellular Matrix Interactions toward Metastasis

doi: 10.1158/0008-5472.can-08-1327

Figure Lengend Snippet: Figure 3. Tumors from wt and SPARC KO mice show similar distribution of epithelial and mesenchymal markers. Immunohistochemical analysis of E-cadherin (A) and N-cadherin (B) in 4T1 tumors from wt and SPARC KO mice. Three different tumor areas from wt and KO mice have been selected to show the distinctive features of E-cadherin expression going from a functional membrane stain (left) to a nonfunctional intracellular (center) or negative stain (right). Tumors from both wt and KO mice show the same pattern of expression of both E-cadherin (A) and N-cadherin (B).

Article Snippet: The underside of the Transwell filter inserts (6.5 mm; Costar) were coated with 10 Ag/mL of fibronectin (Sigma), multimeric vitronectin from bovine plasma (Oxford Biomedical Research, UK), rat tail collagen type I (Becton Dickinson), and/or 200 ng of mouse SPARC purified from parietal yolk sac (Sigma) or recombinant human SPARC (ProSci) overnight at 4jC.

Techniques: Immunohistochemical staining, Expressing, Functional Assay, Membrane, Staining

Journal: Cancer Research

Article Title: Macrophage-Derived SPARC Bridges Tumor Cell-Extracellular Matrix Interactions toward Metastasis

doi: 10.1158/0008-5472.can-08-1327

Figure Lengend Snippet: Figure 4. SPARC modulation of ECM deposition. A, Masson trichrome (a) and silver staining (b) of 4T1 tumors from wt, SPARC KO, and wt > KO chimeras. c, immunohistochemical analysis of fibronectin (FN) in tumors from wt, SPARC KO, and wt > KO chimeras showing a paucity of fibronectin fibers in tumors from SPARC KO mice. B, SPARC synergizes with fibronectin in inducing 4T1 cell migration (B) and such effect is inhibited by mAbs to h1, av, and h5 integrins (C).

Article Snippet: The underside of the Transwell filter inserts (6.5 mm; Costar) were coated with 10 Ag/mL of fibronectin (Sigma), multimeric vitronectin from bovine plasma (Oxford Biomedical Research, UK), rat tail collagen type I (Becton Dickinson), and/or 200 ng of mouse SPARC purified from parietal yolk sac (Sigma) or recombinant human SPARC (ProSci) overnight at 4jC.

Techniques: Silver Staining, Immunohistochemical staining, Migration

Journal: Cancer Research

Article Title: Macrophage-Derived SPARC Bridges Tumor Cell-Extracellular Matrix Interactions toward Metastasis

doi: 10.1158/0008-5472.can-08-1327

Figure Lengend Snippet: Figure 5. SPARC induces 4T1 cell migration. A, migration of PKH-26 labeled 4T1 cells in response to macrophages from wt and SPARC KO mice treated or not with LPS plus IFNg. Migration to both wt and KO macrophages is enhanced by LPS + IFNg but only the increased migration to wt macrophages is inhibited, in part, by a mAb to SPARC (mAb 303). B, dose-dependent SPARC-induced 4T1 cell migration. Different doses of purified murine SPARC were coated onto the underside face of the transwell insert. 4T1 cells were added to the upper chamber of the transwell, and nonmigrated cells were removed after overnight incubation. Migrated cells were counted on the underside of filters with a microscope in 10 randomly chosen fields. Recombinant human SPARC was also tested at the dose in which purified SPARC induces maximal migration. C, effect of integrin inhibitors on SPARC-induced 4T1 migration. 4T1 cells were preincubated with 5 Ag/mL of functional grade endotoxin-free antibodies against integrin av (CD51), integrin h1 (CD29), or SPARC (mAb 303) or mixed with RGD and control RGES peptides. D, SPARC synergizes with multimeric vitronectin (mVN) in promoting 4T1 cell migration. Multimeric vitronectin mixed with SPARC (filled columns) was more active in inducing 4T1 cell migration than either multimeric vitronectin (grey column) or SPARC given alone (empty columns) especially at a low SPARC concentration that mimics a physiologic setting. Columns, mean number of cells per field; bars, SD.

Article Snippet: The underside of the Transwell filter inserts (6.5 mm; Costar) were coated with 10 Ag/mL of fibronectin (Sigma), multimeric vitronectin from bovine plasma (Oxford Biomedical Research, UK), rat tail collagen type I (Becton Dickinson), and/or 200 ng of mouse SPARC purified from parietal yolk sac (Sigma) or recombinant human SPARC (ProSci) overnight at 4jC.

Techniques: Migration, Labeling, Purification, Incubation, Microscopy, Recombinant, Functional Assay, Control, Concentration Assay

Journal: Cancer Research

Article Title: Macrophage-Derived SPARC Bridges Tumor Cell-Extracellular Matrix Interactions toward Metastasis

doi: 10.1158/0008-5472.can-08-1327

Figure Lengend Snippet: Figure 6. 4T1 cells express h5 integrin, which is necessary for cell migration to SPARC and for in vivo metastasis. A, the level of expression of h3 integrin (CD61) and h5 integrin by 4T1 cells cultured in vitro was determined by flow cytometry. Blue, binding of the isotype-matched control antibody; green, target antibody binding. B, expression of h3 and h5 integrins on 4T1 tumor sections. Expression of h3 integrin was confined to cells with endothelial morphology that coexpress CD31 (data not shown). On the contrary, almost all tumor parenchyma stained positive for h5 integrin. Such h5 expression is lost in tumors from 4T1h5 sil cells that still remain negative for h3 integrin. Bar, 100 Am. C, migration of 4T1h5-integrin–stable silenced 4T1 clones (754sil and 1381sil), obtained from two different target sequences, in response to SPARC. Columns, mean number of migrating cells; bars, SD. D, reduced spontaneous lung metastasis by h5 integrin silencing. Stable silenced 754sil

Article Snippet: The underside of the Transwell filter inserts (6.5 mm; Costar) were coated with 10 Ag/mL of fibronectin (Sigma), multimeric vitronectin from bovine plasma (Oxford Biomedical Research, UK), rat tail collagen type I (Becton Dickinson), and/or 200 ng of mouse SPARC purified from parietal yolk sac (Sigma) or recombinant human SPARC (ProSci) overnight at 4jC.

Techniques: Migration, In Vivo, Expressing, Cell Culture, In Vitro, Flow Cytometry, Binding Assay, Control, Staining, Clone Assay