Journal: Journal of Clinical Medicine

Article Title: Expression of PON1, PON2, PON3 and MPO Genes in Patients with Depressive Disorders

doi: 10.3390/jcm11123321

Figure Lengend Snippet: Detailed descriptive statistics for gene (mRNA) expression by study group.

Article Snippet: Reverse Transcription reaction was carried out using a TaqMan ® RNA Reverse Transcription Kit based on the manufacturer’s recommendations, using specific Hs00166557_m1, Hs 00165563_m1, Hs01023629_m1, Hs 99999196_m1 and Hs04194366_g1 probes for PON1, PON2, PON3, MPO and RPL13A genes, respectively, delivered using Applied Biosystems (Foster City, CA, USA).

Techniques: Expressing, Control

Journal: Journal of Clinical Medicine

Article Title: Expression of PON1, PON2, PON3 and MPO Genes in Patients with Depressive Disorders

doi: 10.3390/jcm11123321

Figure Lengend Snippet: Detailed descriptive statistics for protein concentrations (ng/mL) by study group.

Article Snippet: Reverse Transcription reaction was carried out using a TaqMan ® RNA Reverse Transcription Kit based on the manufacturer’s recommendations, using specific Hs00166557_m1, Hs 00165563_m1, Hs01023629_m1, Hs 99999196_m1 and Hs04194366_g1 probes for PON1, PON2, PON3, MPO and RPL13A genes, respectively, delivered using Applied Biosystems (Foster City, CA, USA).

Techniques: Control

Journal: Journal of Clinical Medicine

Article Title: Expression of PON1, PON2, PON3 and MPO Genes in Patients with Depressive Disorders

doi: 10.3390/jcm11123321

Figure Lengend Snippet: Correlation between gene expression and clinical variables. * indicates statistical significance.

Article Snippet: Reverse Transcription reaction was carried out using a TaqMan ® RNA Reverse Transcription Kit based on the manufacturer’s recommendations, using specific Hs00166557_m1, Hs 00165563_m1, Hs01023629_m1, Hs 99999196_m1 and Hs04194366_g1 probes for PON1, PON2, PON3, MPO and RPL13A genes, respectively, delivered using Applied Biosystems (Foster City, CA, USA).

Techniques: Gene Expression

Journal: Journal of Clinical Medicine

Article Title: Expression of PON1, PON2, PON3 and MPO Genes in Patients with Depressive Disorders

doi: 10.3390/jcm11123321

Figure Lengend Snippet: Spearman’s correlations of gene expression with age by study group. * indicates statistical significance.

Article Snippet: Reverse Transcription reaction was carried out using a TaqMan ® RNA Reverse Transcription Kit based on the manufacturer’s recommendations, using specific Hs00166557_m1, Hs 00165563_m1, Hs01023629_m1, Hs 99999196_m1 and Hs04194366_g1 probes for PON1, PON2, PON3, MPO and RPL13A genes, respectively, delivered using Applied Biosystems (Foster City, CA, USA).

Techniques: Gene Expression, Control

Journal: Kidney international

Article Title: Streptococcal pyrogenic exotoxin B antibodies in a mouse model of glomerulonephritis.

doi: 10.1038/sj.ki.5002407

Figure Lengend Snippet: Figure 5 | Leukocyte infiltration in the kidney of mice hyperimmunized with 28-kDa SPE B mutant protein C192S. BALB/ c mice were hyperimmunized four times with (a) BSA or (b) C192S. Leukocyte infiltration was detected using anti-MPO antibody with fluorescence microscopy (n ¼ 10 per group). (c) The mean integral fluorescent intensity of glomeruli was analyzed in 10 fields per mouse (n ¼ 10 mice). Original magnification 200.

Article Snippet: Leukocyte infiltration was detected using anti-MPO antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), and then using FITC-conjugated antigoat IgG (Jackson ImmunoResearch Laboratories Inc.).

Techniques: Mutagenesis, Fluorescence, Microscopy

Journal: Arthritis Research & Therapy

Article Title: Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility

doi: 10.1186/ar4579

Figure Lengend Snippet: RA-derived PMNs exhibited increased spontaneous NETosis and elevated levels of NET-component release. (A) Schematic representation of the time-course design for studying in vitro spontaneous NET release. (B) Detection of in vitro NETosis by immunohistochemistry for neutrophil elastase (NE) (green) and DAPI (blue). Magnification: 20×; scale bar, 50 nm. (C) Concentration of cell-free nucleosomes in PMN culture supernatant by ELISA. (D) Quantification of NET-associated MPO/DNA complexes. These assays indicate that more-rapid and extensive progression of NET formation is observed in RA versus control PMNs. (E) Changes in PMN nuclear morphology during NETosis detected with immunohistochemistry for NE and DAPI. (F) Steady-state (T1) RA-derived PMNs exhibited a greater proportion of delobulated/diffused cells, and progressed rapidly to a NETotic spread phenotype during in vitro culture. Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. All data are representative of at least six independent experiments.

Article Snippet: TaqMan real-time quantitative RT-PCR was performed by using the Applied Biosystems StepOne Plus cycler (Applied Biosystems) and TaqMan Gene Expression Assay primer/probe sets (Applied Biosystems) for ELANE (HS00236952_m1), MPO (HS00924296_m1), PADI4 (HS00202612_m1), and β 2 -microglobulin B2M (HS99999907_m1).

Techniques: Derivative Assay, In Vitro, Immunohistochemistry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Arthritis Research & Therapy

Article Title: Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility

doi: 10.1186/ar4579

Figure Lengend Snippet: Increased expression of NET-associated signaling elements and nuclear localization of PAD4, citrullination of histone H3 in RA-derived PMN, and possible extrusion of PAD4 on NETs. Baseline ROS levels, measured with flow cytometry, were higher in RA-derived PMNs than in control PMNs (A) . Quantitative real-time PCR analysis of NE mRNA (B) and MPO mRNA expression (C) , as well as Western blot analysis of MPO protein levels (D) , indicated that the levels of these two components required for NETosis were elevated in RA-derived PMNs. Total PAD4 protein (E) or PAD4 mRNA expression levels (F) did not indicate any significant difference between control and RA-derived PMNs. (G) Quantification of PAD4 protein levels in cytoplasmic and nuclear fractions of PMNs from healthy controls and RA patients. Nuclear levels of PAD4 were significantly increased in RA patients, whereas the cytoplasmic levels were lower compared with the healthy control PMNs. (H) Elevated citrullinated histone H3 levels in RA PMN extracts detected with Western blot. (I) Co-localization of citrullinated histone H3 (green) with histone components detected with a pan-histone antibody (red), spread over the entire NET surface (blue). Magnification: upper panel 20×, scale bar, 50 nm; lower panel 63× scale bar, 10 nm. (J) Extracellular localization of PAD4 (red) on extruded NETs by multicolor fluorescent immunohistochemistry. NET DNA is stained blue (DAPI), and histones (panH) are stained green. Magnification, 20×; scale bar, 50 nm (K) . Detection of PAD4/cell-free DNA complexes in the culture supernatants of isolated PMN undergoing spontaneous NETosis. Higher levels of these complexes were detected in RA-derived PMN cultures. Time points correspond to those of Figure A. Data are represented as mean ± SEM. ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., statistically not significant. All data are representative of at least six independent experiments.

Article Snippet: TaqMan real-time quantitative RT-PCR was performed by using the Applied Biosystems StepOne Plus cycler (Applied Biosystems) and TaqMan Gene Expression Assay primer/probe sets (Applied Biosystems) for ELANE (HS00236952_m1), MPO (HS00924296_m1), PADI4 (HS00202612_m1), and β 2 -microglobulin B2M (HS99999907_m1).

Techniques: Expressing, Derivative Assay, Flow Cytometry, Control, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Staining, Isolation

Journal: Arthritis Research & Therapy

Article Title: Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility

doi: 10.1186/ar4579

Figure Lengend Snippet: Increased NETotic response of RA-derived PMNs to PMA. (A) Scanning electron micrographs of NETs induced by PMA (25 n M ) indicate the excessive NETotic response of RA-derived PMN. Scale bar, 20 nm. (B) Assessment of NETs induced by PMA treatment by fluorescent immunohistochemistry for MPO (red) and DAPI (blue), indicating the increased response of RA PMNs to PMA (25 n M ). Magnification, 20×; scale bar, 50 nm. (C) Analysis of the nuclear phenotype indicated a vast decrease in delobulated/diffused RA PMN nuclei after treatment with PMA and rapid increase in the NETotic-spread phenotype. (D) Release of cell-free nucleosomes after PMA treatment is abrogated by chloramidine, a PAD4 inhibitor. (E) Increased nuclear localization and concomitant decrease in cytoplasmic PAD4 protein levels after PMA treatment, with a clear tendency for an increased responsiveness to the PMA stimulus by RA PMN. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., statistically not significant. All data are representative of at least four independent experiments.

Article Snippet: TaqMan real-time quantitative RT-PCR was performed by using the Applied Biosystems StepOne Plus cycler (Applied Biosystems) and TaqMan Gene Expression Assay primer/probe sets (Applied Biosystems) for ELANE (HS00236952_m1), MPO (HS00924296_m1), PADI4 (HS00202612_m1), and β 2 -microglobulin B2M (HS99999907_m1).

Techniques: Derivative Assay, Immunohistochemistry

Journal: Arthritis Research & Therapy

Article Title: Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility

doi: 10.1186/ar4579

Figure Lengend Snippet: Influence of RA serum and synovial fluid on normal PMNs. (A) Incubation of healthy donor PMNs with serum (Se) from healthy donors or from RA patients, synovial fluid (SF) from patients with noninflammatory osteoarthritis (OA) or synovial fluid from RA patients. Immunohistochemical analysis of four main components of NETs (NE, MPO, PAD4, and citH3) revealed that RA-derived serum and SF enhanced NETosis in normal PMN compared to healthy control serum or noninflammatory OA SF. PAD4 (white arrowheads) and citH3 (empty arrowheads) colocalize with unmodified histones on NETs. Magnification, 20×; scale bars, 50 nm. (B) Release of cell-free nucleosomes during in vitro culture by PMNs from healthy controls incubated with control serum, RA serum, OA SF or RA SF, or PMA. Data are represented as mean ± SEM. * P < 0.05; ** P < 0.01. (C) Increased ROS generation during in vitro culture by PMNs from healthy controls incubated with control serum, RA serum, OA SF or RA SF, or PMA. Data are presented as mean ± SEM. (D) Release of cell-free nucleosomes during in vitro culture by PMNs from healthy controls, ACPA-positive and ACPA-negative RA patients incubated with serum (Se +), IgG-depleted serum (Se -), or serum reconstituted with their respective eluted IgG, or with PMA. Data are presented as mean ± SEM. * P < 0.05; *** P < 0.001; **** P < 0.0001; n.s., statistically not significant. All data are representative of at least four independent experiments in Figures A–C, and in D.

Article Snippet: TaqMan real-time quantitative RT-PCR was performed by using the Applied Biosystems StepOne Plus cycler (Applied Biosystems) and TaqMan Gene Expression Assay primer/probe sets (Applied Biosystems) for ELANE (HS00236952_m1), MPO (HS00924296_m1), PADI4 (HS00202612_m1), and β 2 -microglobulin B2M (HS99999907_m1).

Techniques: Incubation, Immunohistochemical staining, Derivative Assay, Control, In Vitro

Journal: Arthritis Research & Therapy

Article Title: Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility

doi: 10.1186/ar4579

Figure Lengend Snippet: Elevated serum levels of NET components in RA patients have potential clinical utility. (A) Cell-free DNA levels in plasma and serum from healthy matched blood donors ( n = 41) and patients with RA ( n = 32) determined with real-time PCR. (B) Cell-free nucleosome levels in plasma and serum from healthy donor controls and patients with RA, determined with ELISA. (C) Determination of NE protein concentrations in plasma and serum from healthy donors and patients with RA, as assessed with sandwich ELISA. (D) MPO concentrations in plasma and serum from healthy donors and patients with RA, as determined with sandwich ELISA. (E) NET-associated MPO/DNA complexes quantified by using a modified capture ELISA. In contrast to the serum levels, none of the plasma levels of these NET components attained statistical significance (Figure A to E). (F) ROC analysis of cell-free nucleosomes in serum of patients with RA and healthy controls. (G) Detail of cell-free nucleosome ROC curve with groups of ACPA + and ACPA- RA cases and (H) scatterbox and whisker plots with individual values for control, ACPA + and ACPA- groups. The ROC curve analysis of other NET components, cell-free DNA (I) , NE (J), and MPO (K) , was not as conclusive as that for cell-free nucleosomes. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., statistically not significant.

Article Snippet: TaqMan real-time quantitative RT-PCR was performed by using the Applied Biosystems StepOne Plus cycler (Applied Biosystems) and TaqMan Gene Expression Assay primer/probe sets (Applied Biosystems) for ELANE (HS00236952_m1), MPO (HS00924296_m1), PADI4 (HS00202612_m1), and β 2 -microglobulin B2M (HS99999907_m1).

Techniques: Clinical Proteomics, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Modification, Whisker Assay, Control

Journal: Clinics (Sao Paulo, Brazil)

Article Title: Sex differences in kidney and lung status in an animal model of brain death.

doi: 10.1016/j.clinsp.2025.100623

Figure Lengend Snippet: Fig. 10. Protein expression of MPO and MMP-9 in kidney tissue (red arrow). Naïve, nonmanipulated animals (n = 4); BD, rats subjected to brain death (n = 8). The values represent the means and Standard Errors of the Means (SEMs). Representative photomicrographs (×40) of each group. MPO, Myeloperoxidase; MMP-9, Metalloproteinase-9.

Article Snippet: Kidney markers were MPO (1:100, PA1054 – Boster, USA), MMP-9 (1:100, PB9668 – Boster, USA), eNOS (1:100, AO1604–2 – Boster, USA), iNOS (1:100, AB3523 – Abcam, UK) and caspase-3 (1:100, AB4051 – Abcam, UK).

Techniques: Expressing