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A, B : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from five male healthy donors, (H1 to H5) (A), and five females (H6 to H10) (B) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by <t>densitometry</t> and N-HK concentration was evaluated by the displayed linear regression. C, D : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (C) and HK cleavage (D), measured as in panel A. densito: densitometry in 10 7 arbitrary units; NS: non-significant; ****: p <10 −4 (Mann-Whitney-Wilcoxon test).
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A, B : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from five male healthy donors, (H1 to H5) (A), and five females (H6 to H10) (B) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by <t>densitometry</t> and N-HK concentration was evaluated by the displayed linear regression. C, D : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (C) and HK cleavage (D), measured as in panel A. densito: densitometry in 10 7 arbitrary units; NS: non-significant; ****: p <10 −4 (Mann-Whitney-Wilcoxon test).
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A, B : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from five male healthy donors, (H1 to H5) (A), and five females (H6 to H10) (B) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by <t>densitometry</t> and N-HK concentration was evaluated by the displayed linear regression. C, D : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (C) and HK cleavage (D), measured as in panel A. densito: densitometry in 10 7 arbitrary units; NS: non-significant; ****: p <10 −4 (Mann-Whitney-Wilcoxon test).
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A, B : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from five male healthy donors, (H1 to H5) (A), and five females (H6 to H10) (B) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by densitometry and N-HK concentration was evaluated by the displayed linear regression. C, D : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (C) and HK cleavage (D), measured as in panel A. densito: densitometry in 10 7 arbitrary units; NS: non-significant; ****: p <10 −4 (Mann-Whitney-Wilcoxon test).

Journal: PLoS ONE

Article Title: Kininogen Cleavage Assay: Diagnostic Assistance for Kinin-Mediated Angioedema Conditions

doi: 10.1371/journal.pone.0163958

Figure Lengend Snippet: A, B : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from five male healthy donors, (H1 to H5) (A), and five females (H6 to H10) (B) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by densitometry and N-HK concentration was evaluated by the displayed linear regression. C, D : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (C) and HK cleavage (D), measured as in panel A. densito: densitometry in 10 7 arbitrary units; NS: non-significant; ****: p <10 −4 (Mann-Whitney-Wilcoxon test).

Article Snippet: Using enhanced chemiluminescence (Clarity Western ECL TM substrate, Biorad, Hercules, USA) and densitometry scan quantification (ChemiDoc TM XRS+ System camera and Image Lab TM software, Bio-Rad, Hercules, USA), three molecular species were retained for data processing: native protein (N-HK, 120kDa), light chain (LC-HK, 56kDa), and cleaved light chain (cLC-HK, 46kDa).

Techniques: Purification, Clinical Proteomics, SDS Page, Concentration Assay, MANN-WHITNEY

A : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from nC1Inh-AE male patients (M1 to M5) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by densitometry and N-HK concentration by the displayed linear regression. B, C : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (B) and HK cleavage (C), measured as in panel A, compared to the healthy cohort studied in . D, E, F : as in A, B, C, concerning female patients (W1 to W5). densito: densitometry in 10 7 arbitrary units; ****: p <10 −4 (Mann-Whitney-Wilcoxon test); nC1Inh-AE: angioedema with normal C1 inhibitor.

Journal: PLoS ONE

Article Title: Kininogen Cleavage Assay: Diagnostic Assistance for Kinin-Mediated Angioedema Conditions

doi: 10.1371/journal.pone.0163958

Figure Lengend Snippet: A : Purified human HK (20, 40, 80, and 110ng) and 0.5μL of plasma from nC1Inh-AE male patients (M1 to M5) were subjected to SDS-PAGE. The signal of HK native chain (N-HK), light chain (LC-HK), and cleaved light chain (cLC-HK) were quantified by densitometry and N-HK concentration by the displayed linear regression. B, C : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of plasma concentration of N-HK (B) and HK cleavage (C), measured as in panel A, compared to the healthy cohort studied in . D, E, F : as in A, B, C, concerning female patients (W1 to W5). densito: densitometry in 10 7 arbitrary units; ****: p <10 −4 (Mann-Whitney-Wilcoxon test); nC1Inh-AE: angioedema with normal C1 inhibitor.

Article Snippet: Using enhanced chemiluminescence (Clarity Western ECL TM substrate, Biorad, Hercules, USA) and densitometry scan quantification (ChemiDoc TM XRS+ System camera and Image Lab TM software, Bio-Rad, Hercules, USA), three molecular species were retained for data processing: native protein (N-HK, 120kDa), light chain (LC-HK, 56kDa), and cleaved light chain (cLC-HK, 46kDa).

Techniques: Purification, Clinical Proteomics, SDS Page, Concentration Assay, MANN-WHITNEY

High-molecular-weight kininogen (HK) molecular pattern of plasma samples from healthy women taking or not estrogen/progestin combination (OP; Healthy+OP, n = 32; Healthy-OP, n = 30) and from nC1Inh-AE patients taking OP and ≥2 months after OP withdrawal (nC1INh-AE+/-OP, n = 13). A : Analysis of plasma samples (0.5μL/lane) from six patients (O1–O6) and the corresponding figures. N-HK concentration of P1 and P2 was evaluated by the displayed linear regression, kinin-forming activity was determined as in , pathological values were labelled in bold fonts. B, C : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of the plasma concentration of N-HK (B) and cleaved HK species (C), measured as in panel A. D, E : Details of the paired samples of panel B and C respectively analysed using the Wilcoxon matched-pairs signed rank test. densito: densitometry in 10 7 arbitrary units; NS : non-significant; *: p <0.05; **: p <0.01; ***: p <0.001; ****: p <10 −4 (non-parametric tests); N-HK, LC-HK, cLC-HK: native chain, light chain, cleaved light chain of HK respectively; nC1Inh-AE: angioedema with normal C1 inhibitor.

Journal: PLoS ONE

Article Title: Kininogen Cleavage Assay: Diagnostic Assistance for Kinin-Mediated Angioedema Conditions

doi: 10.1371/journal.pone.0163958

Figure Lengend Snippet: High-molecular-weight kininogen (HK) molecular pattern of plasma samples from healthy women taking or not estrogen/progestin combination (OP; Healthy+OP, n = 32; Healthy-OP, n = 30) and from nC1Inh-AE patients taking OP and ≥2 months after OP withdrawal (nC1INh-AE+/-OP, n = 13). A : Analysis of plasma samples (0.5μL/lane) from six patients (O1–O6) and the corresponding figures. N-HK concentration of P1 and P2 was evaluated by the displayed linear regression, kinin-forming activity was determined as in , pathological values were labelled in bold fonts. B, C : Box-plot displaying 5–95% range ( italic fonts), median ( bold fonts), and interquartile values of the plasma concentration of N-HK (B) and cleaved HK species (C), measured as in panel A. D, E : Details of the paired samples of panel B and C respectively analysed using the Wilcoxon matched-pairs signed rank test. densito: densitometry in 10 7 arbitrary units; NS : non-significant; *: p <0.05; **: p <0.01; ***: p <0.001; ****: p <10 −4 (non-parametric tests); N-HK, LC-HK, cLC-HK: native chain, light chain, cleaved light chain of HK respectively; nC1Inh-AE: angioedema with normal C1 inhibitor.

Article Snippet: Using enhanced chemiluminescence (Clarity Western ECL TM substrate, Biorad, Hercules, USA) and densitometry scan quantification (ChemiDoc TM XRS+ System camera and Image Lab TM software, Bio-Rad, Hercules, USA), three molecular species were retained for data processing: native protein (N-HK, 120kDa), light chain (LC-HK, 56kDa), and cleaved light chain (cLC-HK, 46kDa).

Techniques: High Molecular Weight, Clinical Proteomics, Concentration Assay, Activity Assay

A : High-molecular-weight kininogen (HK) molecular pattern of plasma samples (0.5μL/lane) from two nC1Inh-AE patients (W6, M6) less than 24h after the onset of an AE attack (Att) and at least two weeks after remission (Rem), analysed as in . B : Same as for A, with plasma samples from three patients (K1–K3) with ACEi-AE and low kinin catabolism (L-Cat) and three patients (M7–M9) with nC1Inh-AE and normal kinin catabolism (N-Cat). N-HK concentrations were evaluated by the displayed linear regressions, kinin-forming activity was determined as in , pathological values were labelled in bold font. densito: densitometry in 10 7 arbitrary units; N-HK, LC-HK, cLC-HK: native chain, light chain, cleaved light chain of HK respectively.

Journal: PLoS ONE

Article Title: Kininogen Cleavage Assay: Diagnostic Assistance for Kinin-Mediated Angioedema Conditions

doi: 10.1371/journal.pone.0163958

Figure Lengend Snippet: A : High-molecular-weight kininogen (HK) molecular pattern of plasma samples (0.5μL/lane) from two nC1Inh-AE patients (W6, M6) less than 24h after the onset of an AE attack (Att) and at least two weeks after remission (Rem), analysed as in . B : Same as for A, with plasma samples from three patients (K1–K3) with ACEi-AE and low kinin catabolism (L-Cat) and three patients (M7–M9) with nC1Inh-AE and normal kinin catabolism (N-Cat). N-HK concentrations were evaluated by the displayed linear regressions, kinin-forming activity was determined as in , pathological values were labelled in bold font. densito: densitometry in 10 7 arbitrary units; N-HK, LC-HK, cLC-HK: native chain, light chain, cleaved light chain of HK respectively.

Article Snippet: Using enhanced chemiluminescence (Clarity Western ECL TM substrate, Biorad, Hercules, USA) and densitometry scan quantification (ChemiDoc TM XRS+ System camera and Image Lab TM software, Bio-Rad, Hercules, USA), three molecular species were retained for data processing: native protein (N-HK, 120kDa), light chain (LC-HK, 56kDa), and cleaved light chain (cLC-HK, 46kDa).

Techniques: High Molecular Weight, Clinical Proteomics, Activity Assay