Journal: The Journal of rheumatology

Article Title: Neutrophil Extracellular Traps (NETs) profiles in patients with incident SLE and lupus nephritis

doi: 10.3899/jrheum.181232

Figure Lengend Snippet: (A) DNASE1 serum levels. A homemade ELISA assay has been utilized to test DNASE1. Results are expressed as ng/ml and represented as median and interquartile range.

Article Snippet: The assay was validated by comparison with a commercial kit for DNase1 (Catalog n LS-F27682 LSBio Inc., Seattle, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Regenerative Biomaterials

Article Title: In vitro evaluation of 3D printed polycaprolactone scaffolds with angle-ply architecture for annulus fibrosus tissue engineering

doi: 10.1093/rb/rbz011

Figure Lengend Snippet: Immunocytochemical results at Day 21 of culture for bovine AF cells seeded on smooth PCL surfaces. Scaffolds were treated with primary antibodies targeting ( a ) ACAN, ( b ) COL 1, ( c ) COL 2, ( d ) TNMD. Alexa Fluor ® 647 was used for secondary antibody, and cells were counterstained with ethidium bromide. Cell nuclei are shown in cyan to enhance contrast. Scale bars=100 μm

Article Snippet: Scaffolds were incubated at room temperature for 1 h in mouse monoclonal antibody against bovine collagen type I (COL1, Abcam, 1:100 dilution), collagen type II (COL2, Abcam, 1:200 dilution), aggrecan (ACAN, Abcam, 1:50 dilution) or rabbit anti-bovine polyclonal antibody against tenomodulin (TNMD, Fisher Scientific, 1:100 dilution).

Techniques:

Journal: Regenerative Biomaterials

Article Title: In vitro evaluation of 3D printed polycaprolactone scaffolds with angle-ply architecture for annulus fibrosus tissue engineering

doi: 10.1093/rb/rbz011

Figure Lengend Snippet: Immunocytochemical results at Day 21 of culture for bovine AF cells seeded on smooth PCL surfaces. Scaffolds were treated with primary antibodies targeting ( a ) ACAN, ( b ) COL 1, ( c ) COL 2, ( d ) TNMD. Alexa Fluor ® 647 was used for secondary antibody and cells were counterstained with ethidium bromide. Cell nuclei are shown in cyan to enhance contrast. Scale bars=100 μm

Article Snippet: Scaffolds were incubated at room temperature for 1 h in mouse monoclonal antibody against bovine collagen type I (COL1, Abcam, 1:100 dilution), collagen type II (COL2, Abcam, 1:200 dilution), aggrecan (ACAN, Abcam, 1:50 dilution) or rabbit anti-bovine polyclonal antibody against tenomodulin (TNMD, Fisher Scientific, 1:100 dilution).

Techniques:

Journal: Frontiers in Immunology

Article Title: Immunomodulation of Host Chitinase 3-Like 1 During a Mammary Pathogenic Escherichia coli Infection

doi: 10.3389/fimmu.2018.01143

Figure Lengend Snippet: Chitinase 3-like 1 (CHI3L1) in coliform-infected bovine samples. (A) Bovine CHI3L1 protein detection in whey isolated from naturally coliform-infected cows ( n = 6) compared to whey isolated from healthy quarters ( n = 3). (B) interleukin (IL)-1beta, IL-6, IL-8, and CHI3L1 expression in Escherichia coli 1303 experimentally infected cows ( n ≥ 4). Data are presented as means ± SEM, indicating significant differences with *( P < 0.05).

Article Snippet: To detect bovine CHI3L1, the membranes were blocked in Tris-Buffered Saline (TBS) supplemented with 0.1% Tween-20 (v/v), 0.1% bovine serum albumin, and 4% (w/v) non-fat dry milk and incubated overnight at 2–8°C on a platform shaker with a rabbit anti-bovine CHI3L1 polyclonal/monoclonal antibody (1.5 mg ml −1 ), kindly provided by Tom Wheeler, AgResearch, diluted 1:1,000 in TBS supplemented as described ( ).

Techniques: Infection, Isolation, Expressing

Journal: Frontiers in Immunology

Article Title: Immunomodulation of Host Chitinase 3-Like 1 During a Mammary Pathogenic Escherichia coli Infection

doi: 10.3389/fimmu.2018.01143

Figure Lengend Snippet: Mammary chitinase 3-like 1 (CHI3L1) during mammary pathogenic E. coli (MPEC) and non-MPEC infections in the mouse mastitis model. (A) Bacterial loads in the mammary gland at 24 h p.i. with MPEC strains Escherichia coli P4 and 1303 compared to non-MPEC strains E. coli K12 strains DH5alpha and MG1655, and phosphate buffered saline (PBS) controls ( n ≥ 8). (B) Histology of mammary tissue at 24 h p.i. with MPEC strains E. coli P4 and 1303, and non-MPEC strains E. coli K12 strains DH5alpha and MG1655 compared to PBS controls. (C) Association of IL-8 levels and bacterial loads in mammary tissue at 24 h p.i. with MPEC strains E. coli P4 and 1303, and non-MPEC strains E. coli K12 strains DH5alpha and MG1655 ( n ≥ 7; r = 0.63; P < 0.01; 95% C.I. 0.354–0.808). (D) CHI3L1 levels in protein lysates extracted from mammary tissue at 24 h p.i. with MPEC strains E. coli P4 and 1303, and non-MPEC strains E. coli K12 strains DH5alpha and MG1655 compared to PBS controls ( n ≥ 8). Data are presented as means ± SEM, indicating significant differences with *( P < 0.05); **( P < 0.01) or ***( P < 0.001). ns, not significant.

Article Snippet: To detect bovine CHI3L1, the membranes were blocked in Tris-Buffered Saline (TBS) supplemented with 0.1% Tween-20 (v/v), 0.1% bovine serum albumin, and 4% (w/v) non-fat dry milk and incubated overnight at 2–8°C on a platform shaker with a rabbit anti-bovine CHI3L1 polyclonal/monoclonal antibody (1.5 mg ml −1 ), kindly provided by Tom Wheeler, AgResearch, diluted 1:1,000 in TBS supplemented as described ( ).

Techniques: Saline

Journal: Frontiers in Immunology

Article Title: Immunomodulation of Host Chitinase 3-Like 1 During a Mammary Pathogenic Escherichia coli Infection

doi: 10.3389/fimmu.2018.01143

Figure Lengend Snippet: Downstream effects of mammary chitinase 3-like 1 (CHI3L1) during Escherichia coli P4 infections in the mouse mastitis model. (A) Bacterial loads ( n ≥ 12), and interleukin (IL)-8 ( n ≥ 3), CHI3L1 ( n ≥ 5), and RANTES/CCL5 levels ( n ≥ 3) in the mammary glands of CHI3L1 +/+ and CHI3L1 −/− mice 24 h p.i. with E. coli P4. (B) IL-1beta ( n ≥ 3) and IL-6 levels ( n ≥ 3) in the mammary gland of CHI3L1 +/+ and CHI3L1 −/− mice 24 h p.i. with E. coli P4. (C) H&E histology of mammary tissue 24 h p.i. with E. coli P4 in CHI3L1 +/+ and CHI3L1 −/− mice. Black arrow illustrates the presumed neutrophilic innate immune cells (based on their characteristic polymorphonuclear morphology) in the interstitial space. (D) Immunohistochemical stainings for Ly6G on paraffin slides of mammary glands at 24 h p.i. with E. coli P4 or phosphate buffered saline (PBS) in CHI3L1 +/+ and CHI3L1 −/− mice. Black arrows indicate Ly6G-positive neutrophils in the interstitial space. Scale bars = 20 µm. The bar graphs display the quantified percentage of Ly6G-positive cells in the interstitial space versus the luminal space for stained sections of E. coli P4-infected mammary glands from CHI3L1 +/+ and CHI3L1 −/− mice ( n ≥ 2 glands). Data are presented as means ± SEM, indicating significant differences with **( P < 0.01) or ***( P < 0.001). ns, not significant.

Article Snippet: To detect bovine CHI3L1, the membranes were blocked in Tris-Buffered Saline (TBS) supplemented with 0.1% Tween-20 (v/v), 0.1% bovine serum albumin, and 4% (w/v) non-fat dry milk and incubated overnight at 2–8°C on a platform shaker with a rabbit anti-bovine CHI3L1 polyclonal/monoclonal antibody (1.5 mg ml −1 ), kindly provided by Tom Wheeler, AgResearch, diluted 1:1,000 in TBS supplemented as described ( ).

Techniques: Immunohistochemical staining, Saline, Staining, Infection

Journal: Frontiers in Immunology

Article Title: Immunomodulation of Host Chitinase 3-Like 1 During a Mammary Pathogenic Escherichia coli Infection

doi: 10.3389/fimmu.2018.01143

Figure Lengend Snippet: Upstream regulators of mammary chitinase 3-like 1 (CHI3L1) during Escherichia coli P4 infections in the mouse mastitis model. (A) Bacterial loads ( n = 5), and interleukin (IL)-8 ( n = 5), CHI3L1 ( n ≥ 4), and RANTES/CCL5 levels ( n = 5) in the E. coli P4-infected mammary glands of mice treated with Z-VAD-fmk or DMSO (sham) at 24 h p.i. (B) Bacterial loads ( n ≥ 12), and IL-8 ( n ≥ 8), CHI3L1 ( n = 6), and RANTES/CCL5 levels ( n = 6) in the mammary gland of mice treated with chitin or phosphate buffered saline (PBS) controls at 24 h p.i. with E. coli P4. (C) IL-17A ( n = 6), IL-4 ( n = 6), and MCP-1 levels ( n = 6) in the E. coli P4-infected mammary gland of mice treated with chitin or PBS controls at 24 h p.i. Data are presented as means ± SEM, indicating significant differences with *( P < 0.05); **( P < 0.01); or ***( P < 0.001). ns, not significant.

Article Snippet: To detect bovine CHI3L1, the membranes were blocked in Tris-Buffered Saline (TBS) supplemented with 0.1% Tween-20 (v/v), 0.1% bovine serum albumin, and 4% (w/v) non-fat dry milk and incubated overnight at 2–8°C on a platform shaker with a rabbit anti-bovine CHI3L1 polyclonal/monoclonal antibody (1.5 mg ml −1 ), kindly provided by Tom Wheeler, AgResearch, diluted 1:1,000 in TBS supplemented as described ( ).

Techniques: Infection, Saline

Journal: The Journal of Reproduction and Development

Article Title: Expression of C–C motif chemokines and their receptors in bovine placentomes at spontaneous and induced parturition

doi: 10.1262/jrd.2019-113

Figure Lengend Snippet: Transcripts upregulated and downregulated in caruncle tissue at spontaneous parturition compared with induced (DEX group) parturition

Article Snippet: The 5 µm-thick sections from formalin-fixed and paraffin-embedded tissue were incubated at room temperature with rabbit polyclonal anti-bovine CCL8 antibody (MBS2026335; 1:400; MyBioSource, San Diego, CA), rabbit polyclonal anti-human CCR1 antibody (ab140756; 1:200; Abcam PLC, Cambridge, UK), rabbit polyclonal anti-human CCR2 antibody (NBP1-48337; 1:100; Novus Biologicals LLC, Littleton, CO), or rabbit polyclonal anti-human CCR5 antibody (NBP2-31374SS; 1:100; Novus Biologicals LLC) for 12 h. The signals were detected using anti-rabbit IgG-biotin conjugate (Sigma-Aldrich, St. Louis, MO, USA) diluted 1:100 for 1 h and then counterstained with hematoxylin.

Techniques:

Journal: The Journal of Reproduction and Development

Article Title: Expression of C–C motif chemokines and their receptors in bovine placentomes at spontaneous and induced parturition

doi: 10.1262/jrd.2019-113

Figure Lengend Snippet: mRNA expression levels of CCLs and their receptors in the placentome at spontaneous (SP group) and induced (PG, DEX, and TABET groups) parturition. A, CCL2 ; B, CCL8 ; C, CCR1 ; and D, CCR5 mRNA expression. Statistically significant differences in the relative abundance of CCL2 , CCL8 , CCR1 , and CCR5 mRNA were analyzed in cotyledon (COT) and caruncle (CAR) tissue. Data are presented as the mean ± standard error. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The 5 µm-thick sections from formalin-fixed and paraffin-embedded tissue were incubated at room temperature with rabbit polyclonal anti-bovine CCL8 antibody (MBS2026335; 1:400; MyBioSource, San Diego, CA), rabbit polyclonal anti-human CCR1 antibody (ab140756; 1:200; Abcam PLC, Cambridge, UK), rabbit polyclonal anti-human CCR2 antibody (NBP1-48337; 1:100; Novus Biologicals LLC, Littleton, CO), or rabbit polyclonal anti-human CCR5 antibody (NBP2-31374SS; 1:100; Novus Biologicals LLC) for 12 h. The signals were detected using anti-rabbit IgG-biotin conjugate (Sigma-Aldrich, St. Louis, MO, USA) diluted 1:100 for 1 h and then counterstained with hematoxylin.

Techniques: Expressing

Journal: The Journal of Reproduction and Development

Article Title: Expression of C–C motif chemokines and their receptors in bovine placentomes at spontaneous and induced parturition

doi: 10.1262/jrd.2019-113

Figure Lengend Snippet: Primer sequences for PCR

Article Snippet: The 5 µm-thick sections from formalin-fixed and paraffin-embedded tissue were incubated at room temperature with rabbit polyclonal anti-bovine CCL8 antibody (MBS2026335; 1:400; MyBioSource, San Diego, CA), rabbit polyclonal anti-human CCR1 antibody (ab140756; 1:200; Abcam PLC, Cambridge, UK), rabbit polyclonal anti-human CCR2 antibody (NBP1-48337; 1:100; Novus Biologicals LLC, Littleton, CO), or rabbit polyclonal anti-human CCR5 antibody (NBP2-31374SS; 1:100; Novus Biologicals LLC) for 12 h. The signals were detected using anti-rabbit IgG-biotin conjugate (Sigma-Aldrich, St. Louis, MO, USA) diluted 1:100 for 1 h and then counterstained with hematoxylin.

Techniques: Sequencing