Journal: International Journal of Molecular Sciences

Article Title: The Dual Role of Neutrophil Extracellular Traps (NETs) in Sepsis and Ischemia-Reperfusion Injury: Comparative Analysis across Murine Models

doi: 10.3390/ijms25073787

Figure Lengend Snippet: NETs differentially modulate the 24 h survival rate in sepsis induced by volvulus, LPS or CLP. ( A ) Survival rates for the volvulus model for WT ( n = 8, 62.5%), PAD4 -KO ( n = 8, 75%), and DNase1/1L3 -DKO ( n = 8, 50%) groups at 48 h. ( B ) Survival rate for the LPS model for WT ( n = 7, 71.4%), PAD4 -KO ( n = 8, 87.5%), and DNase1/1L3 -DKO ( n = 8, 37.5%) groups at 48 h. ( C ) Survival rates for the CLP model for WT ( n = 11, 45.4%), PAD4 -KO ( n = 8, 25%), and DNase1/1L3 -DKO ( n = 8, 62.5%) groups at 48 h. The Mantel–Cox (log-rank) test was applied to compare the survival distributions among the groups. p -values are indicated above, showing that the findings were not significant.

Article Snippet: Sections were incubated overnight at 4 °C with DNase1-polyclonal antibody (BS-7651R, Bioss, Woburn, MA, USA, 0.01 µg/mL) or DNase1L3-polyclonal antibody (BS-7653R, Bioss, Woburn, MA, USA, 0.01 µg/mL).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Dual Role of Neutrophil Extracellular Traps (NETs) in Sepsis and Ischemia-Reperfusion Injury: Comparative Analysis across Murine Models

doi: 10.3390/ijms25073787

Figure Lengend Snippet: During sepsis, cfDNA, DNase1, and neutrophil elastase (NE) plasma levels increase. Plasma levels of the canonical NET markers cfDNA, DNase, and NE are increased in all three sepsis models. ( A – C ) The levels of cfDNA in volvulus ( A ), LPS ( B ), and CLP ( C ); ( D – F ) levels of DNase in volvulus ( D ), LPS ( E ), and CLP ( F ); NE plasma levels in volvulus ( G ), LPS ( H ), and CLP ( I ) 48 h after intervention. Ordinary one-way ANOVA (Analysis of Variance) was used. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Sections were incubated overnight at 4 °C with DNase1-polyclonal antibody (BS-7651R, Bioss, Woburn, MA, USA, 0.01 µg/mL) or DNase1L3-polyclonal antibody (BS-7653R, Bioss, Woburn, MA, USA, 0.01 µg/mL).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Dual Role of Neutrophil Extracellular Traps (NETs) in Sepsis and Ischemia-Reperfusion Injury: Comparative Analysis across Murine Models

doi: 10.3390/ijms25073787

Figure Lengend Snippet: DNase1 and Dnase1L3 expression is increased in PAD4 -KO mice. Representative merged images of immunofluorescence staining for DNase1 and DNase1L3 in the intestine, liver, and lung: ( A ) intestine, 40× magnification; ( B ) liver, 40× magnification; ( C ) lung, 40× magnification; ( D ) control: median laparotomy, exposure of the intestine for 1 min, closing the abdomen, no further manipulation. Magnification was 40×. DNA was stained with DAPI (blue). treat, treatment groups.

Article Snippet: Sections were incubated overnight at 4 °C with DNase1-polyclonal antibody (BS-7651R, Bioss, Woburn, MA, USA, 0.01 µg/mL) or DNase1L3-polyclonal antibody (BS-7653R, Bioss, Woburn, MA, USA, 0.01 µg/mL).

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: International Journal of Molecular Sciences

Article Title: The Dual Role of Neutrophil Extracellular Traps (NETs) in Sepsis and Ischemia-Reperfusion Injury: Comparative Analysis across Murine Models

doi: 10.3390/ijms25073787

Figure Lengend Snippet: PAD4 -KO and DNase1/1L3 -DKO mice show reduced oxidative stress. MDA levels in liver, lung, and spleen ( A , D , G ) of volvulus, ( B , E , H ) LPS, and ( C , F , I ) CLP-induced sepsis, respectively. Ordinary one-way ANOVA was used. ns, not significant. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Sections were incubated overnight at 4 °C with DNase1-polyclonal antibody (BS-7651R, Bioss, Woburn, MA, USA, 0.01 µg/mL) or DNase1L3-polyclonal antibody (BS-7653R, Bioss, Woburn, MA, USA, 0.01 µg/mL).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Dual Role of Neutrophil Extracellular Traps (NETs) in Sepsis and Ischemia-Reperfusion Injury: Comparative Analysis across Murine Models

doi: 10.3390/ijms25073787

Figure Lengend Snippet: The antioxidant capacity is impaired in DNase1/1L3 -DKO mice. GPx levels in lungs, intestines, and livers of ( A , D , G ) volvulus, ( B , E , H ) LPS, and ( C , F , I ) CLP-induced sepsis, respectively. Ordinary one-way ANOVA was used. ns, not significant * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Sections were incubated overnight at 4 °C with DNase1-polyclonal antibody (BS-7651R, Bioss, Woburn, MA, USA, 0.01 µg/mL) or DNase1L3-polyclonal antibody (BS-7653R, Bioss, Woburn, MA, USA, 0.01 µg/mL).

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Soluble Urokinase Plasminogen Activator Receptor as a Predictor of All-Cause Death in Patients Undergoing Coronary Angiography at 10-Year Follow-Up

doi: 10.3390/jcm13206158

Figure Lengend Snippet: Biochemical tests based on suPAR concentration.

Article Snippet: Deoxyribonuclease 1 (DNase1) was measured using the Human DNASE1 ELISA Kit (Novus Biologicals, a Bio-Techne brand, Abingdon, UK), and neutrophil elastase was quantified using the Human Neutrophil Elastase SimpleStep ELISA Kit (Abcam, Cambridge, UK), following similar procedures to those described above.

Techniques: Concentration Assay

Journal: Molecular and Cellular Endocrinology

Article Title: Exposure to chemical cocktails before or after conception – The effect of timing on ovarian development ☆

doi: 10.1016/j.mce.2013.06.016

Figure Lengend Snippet: Fetal ovarian proteins exhibiting differential expression following continuous or cross-over exposure to chemical cocktails in sewage sludge fertiliser. The accession number is derived from NCBI. Fold change relative to the normalised spot volumes for the CC group are increased if marked “+” and decreased if marked “−“. Fold-change values attaining statistical significance are highlighted in bold. P values are derived by post hoc tests of log-normalised spot volumes. Spots containing significant protein matches that cannot be discriminated between are marked with * next to the spot number. Where the same protein is identified in different spots vertical lines join the spot numbers.

Article Snippet: Antibodies used were: (i) HSP60 1:25, mouse (ab1819, Abcam); (ii) HSP70 1:800, mouse (ab47455, Abcam); (iii) HSP90 1:500, mouse (H1775, Sigma–Aldrich); (iv) HSPA4L 2.5 μg/ml, goat (LS-B2621, Life Span Biosciences Inc.); (v) HSF1 1:75, rabbit (ab47484, Abcam); (vi) MVP 1:200, mouse (ab14562, Abcam); (vii) DNASE1 1:1000, rabbit (18-732-292145 GenWay); (viii) ANXA1 1:125, goat (LS-B3028/26040, Life Span Biosciences Inc.); (ix) CDKN1B 1:100, rabbit (GTX27961, GeneTex Inc.); (x) IDH1 1:100, rabbit (ab36329, Abcam); (xi) GSTM3 1:100, rabbit (15214-1-AP, ProteinTech) which were incubated for 30 min at room temperature.

Techniques: Quantitative Proteomics, Derivative Assay, Activity Assay, Binding Assay, Membrane, Transduction, Conjugation Assay

Journal: Molecular and Cellular Endocrinology

Article Title: Exposure to chemical cocktails before or after conception – The effect of timing on ovarian development ☆

doi: 10.1016/j.mce.2013.06.016

Figure Lengend Snippet: Networks affected by sewage sludge exposure. Genes shown in bold are those, or their products, identified as significantly altered in at least one treatment group compared with controls (CC) in the present study. Analysis was performed using IPA.

Article Snippet: Antibodies used were: (i) HSP60 1:25, mouse (ab1819, Abcam); (ii) HSP70 1:800, mouse (ab47455, Abcam); (iii) HSP90 1:500, mouse (H1775, Sigma–Aldrich); (iv) HSPA4L 2.5 μg/ml, goat (LS-B2621, Life Span Biosciences Inc.); (v) HSF1 1:75, rabbit (ab47484, Abcam); (vi) MVP 1:200, mouse (ab14562, Abcam); (vii) DNASE1 1:1000, rabbit (18-732-292145 GenWay); (viii) ANXA1 1:125, goat (LS-B3028/26040, Life Span Biosciences Inc.); (ix) CDKN1B 1:100, rabbit (GTX27961, GeneTex Inc.); (x) IDH1 1:100, rabbit (ab36329, Abcam); (xi) GSTM3 1:100, rabbit (15214-1-AP, ProteinTech) which were incubated for 30 min at room temperature.

Techniques: Ubiquitin Proteomics, Modification

Journal: Molecular and Cellular Endocrinology

Article Title: Exposure to chemical cocktails before or after conception – The effect of timing on ovarian development ☆

doi: 10.1016/j.mce.2013.06.016

Figure Lengend Snippet: Immunolocalisation of proteins identified as affected by sewage sludge exposure in the fetal sheep ovary. Heat-shock proteins, HSP60 (A), HSP70 (B), HSP90 (C) and HSPA4L (D) were all predominantly oocyte-specific, with strong cytoplasmic staining in most oocytes (blue arrows) but not all oocytes (white arrows). The fifth heat-shock protein, HSF1 (E) was localised in granulosa cells, pre-granulosa cells and many (but not all) somatic cells around the follicles. At higher power, HSF1 expression was detected in the nuclei of granulosa cells (blue arrows), but not in all pre-granulosa cells around primordial or forming primordial follicles (white arrow). CDKN1B (F) was localised in the cytoplasm and/or nuclei of some but not all oocytes and in the nuclei of some granulosa cells (blue arrows). Atretic follicles were CDKN1B-negative, but so were some healthy oocytes and follicles (white arrows). DNASE1 (G) showed intense staining in somatic cells around blood vessels and mesenchymal remnants (blue arrow) although all oocytes were negative (white arrow). ANXA1 (H) exhibited quite wide-spread staining, especially in the ovarian surface epithelium, the cytoplasm of many oocytes and scattered somatic cells (blue arrows). GSTM3 (I) was also principally localised to oocyte cytoplasm (blue arrow) although some oocytes, particularly those showing signs of atresia or dark condensed nuclei were negative (white arrow). IDH1 (J) was localised mainly in oocyte cytoplasm (blue arrow) but also in some somatic cells, including some surface epithelium cells. Positive staining is brown (DAB), counterstained by haematoxylin (blue). The bars denote scale for each image separately. Blue arrows highlight immuno-positive cells and white arrows immuno-negative cells. Two magnifications, taken from different ovaries, are shown for each antigen, separated by a white line. Each antigen is bounded by a black box to simplify interpretation. In all cases IgG-negative slides incubated with non-immune serum of the appropriate species were characterised by an absence of brown stain (one inset panel for each antigen).

Article Snippet: Antibodies used were: (i) HSP60 1:25, mouse (ab1819, Abcam); (ii) HSP70 1:800, mouse (ab47455, Abcam); (iii) HSP90 1:500, mouse (H1775, Sigma–Aldrich); (iv) HSPA4L 2.5 μg/ml, goat (LS-B2621, Life Span Biosciences Inc.); (v) HSF1 1:75, rabbit (ab47484, Abcam); (vi) MVP 1:200, mouse (ab14562, Abcam); (vii) DNASE1 1:1000, rabbit (18-732-292145 GenWay); (viii) ANXA1 1:125, goat (LS-B3028/26040, Life Span Biosciences Inc.); (ix) CDKN1B 1:100, rabbit (GTX27961, GeneTex Inc.); (x) IDH1 1:100, rabbit (ab36329, Abcam); (xi) GSTM3 1:100, rabbit (15214-1-AP, ProteinTech) which were incubated for 30 min at room temperature.

Techniques: Staining, Expressing, Incubation

Journal: Journal of Clinical Medicine

Article Title: Soluble Urokinase Plasminogen Activator Receptor as a Predictor of All-Cause Death in Patients Undergoing Coronary Angiography at 10-Year Follow-Up

doi: 10.3390/jcm13206158

Figure Lengend Snippet: Biochemical tests based on suPAR concentration.

Article Snippet: Deoxyribonuclease 1 (DNase1) was measured using the Human DNASE1 ELISA Kit (Novus Biologicals, a Bio-Techne brand, Abingdon, UK), and neutrophil elastase was quantified using the Human Neutrophil Elastase SimpleStep ELISA Kit (Abcam, Cambridge, UK), following similar procedures to those described above.

Techniques: Concentration Assay