Journal: iScience

Article Title: Dietary fiber reduces mortality from secondary sepsis in a murine model of Clostridioides difficile infection

doi: 10.1016/j.isci.2026.115258

Figure Lengend Snippet: Plasma markers of systemic inflammation and kidney injury in CDI mice across diets (A) PCoA of Canberra distances of mean-normalized sepsis/immune marker concentrations. Points are colored by diet, and shapes indicate whether mice were humanely euthanized due to clinical sickness or survived until the experimental endpoint. Vectors show the correlation of each measured factor with PC1 and PC2, with the vector length indicating the relative strength of the correlation. Only statistically significant vectors are shown (multiple regression with Benjamini-Hochberg multiple test correction, p.adj. < 0.05). (B) Plasma concentration of each marker that significantly differed between diets (blood urea nitrogen (BUN), and immune factors sCD14, CXCL1, IL-10, IL-1B, IL-6, and TNF-a) at sacrifice. Pairwise comparisons of concentrations between diets were calculated using Kruskal-Wallis and Dunn’s post hoc tests, with p -value corrections conducted via Benjamini and Hochberg. Boxplot lines (from top to bottom) depict the 75 th , 50 th (median), and 25 th percentiles, with lines extending from the top/bottom of the boxplot indicating the largest/smallest observation within ±1.5∗IQR (inter-quartile range). P-value significance (∗∗∗∗: p < 0.0001, ∗∗∗: p < 0.001, ∗∗: p < 0.01, and ∗: p < 0.05).

Article Snippet: Soluble CD14 (sCD14) levels were quantified using the Quantikine Mouse sCD14 ELISA Kit (Catalog No. MC140, R&D Systems, Minneapolis, MN, USA) following the manufacturer’s instructions.

Techniques: Clinical Proteomics, Marker, Plasmid Preparation, Concentration Assay

Journal: Scientific reports

Article Title: Blood-brain barrier and gut barrier dysfunction in chronic kidney disease with a focus on circulating biomarkers and tight junction proteins.

doi: 10.1038/s41598-022-08387-7

Figure Lengend Snippet: Figure 2. Analysis of soluble CD14 (sCD14) levels in haemodialysis (HD) and non-CKD controls (control). (A) Soluble CD14 levels. The number of samples (n) that each analyte was detected in is noted for each group. Data presented at median and interquartile range. Statistical significance, ***p < 0.001, ****p < 0.0001. (B) Spearman’s correlation between sCD14 and trimethylamine N-oxide (TMAO) levels.

Article Snippet: The S100B ELISA kit was performed in accordance with manufacturer guidelines, with the alteration that samples were undiluted. sCD14 levels in HD patients was previously recorded and reported87, for measurements in non-CKD controls the same CD14 ELISA kit (DC140, R&D Systems, UK) was used, and run in accordance with manufacturer guidelines.

Techniques: Control

Journal: Scientific reports

Article Title: Blood-brain barrier and gut barrier dysfunction in chronic kidney disease with a focus on circulating biomarkers and tight junction proteins.

doi: 10.1038/s41598-022-08387-7

Figure Lengend Snippet: Figure 1. Analysis of serum biomarkers in haemodialysis (HD) and non-CKD controls (control). (A) Brain-derived neurotrophic factor (BDNF), (B) Neuron-specific enolase (NSE) were measured by enzyme- linked immunosorbent assay (ELISA). (C) Trimethylamine N-oxide (TMAO) was measured by liquid chromatography-mass spectrometry (LC–MS). The number of samples (n) that each analyte was detected in is noted for each group. Data presented at median and interquartile range. Statistical significance, ***p < 0.001, ****p < 0.0001. (D) Spearman’s correlation between BDNF and TMAO levels. (E) Spearman’s correlation between NSE and TMAO levels.

Article Snippet: The S100B ELISA kit was performed in accordance with manufacturer guidelines, with the alteration that samples were undiluted. sCD14 levels in HD patients was previously recorded and reported87, for measurements in non-CKD controls the same CD14 ELISA kit (DC140, R&D Systems, UK) was used, and run in accordance with manufacturer guidelines.

Techniques: Control, Derivative Assay, Enzyme-linked Immunosorbent Assay, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy

Journal: Annals of Gastroenterology : Quarterly Publication of the Hellenic Society of Gastroenterology

Article Title: Systemic levels of human β-defensin 1 are elevated in patients with cirrhosis

doi:

Figure Lengend Snippet: Elevated soluble CD14 (sCD14) concentration in the serum of cirrhotic patients. Concentration of sCD14 was measured in healthy controls (n=31), patients with chronic viral hepatitis (n=26) and patients with cirrhosis (n=50) as described in patients and methods. Data is presented graphically as Whisker box-plots

Article Snippet: For sCD14, human CD14 ELISA Development kit (R&D Systems, Abingdon, UK) was used following manufacturer’s instructions.

Techniques: Concentration Assay, Whisker Assay

Journal: Annals of Gastroenterology : Quarterly Publication of the Hellenic Society of Gastroenterology

Article Title: Systemic levels of human β-defensin 1 are elevated in patients with cirrhosis

doi:

Figure Lengend Snippet: High correlation between the levels of human beta defensin-1 (hBD-1) and soluble CD14 (sCD14) in the hepatic veins of cirrhotic patients . Concentrations of hBD-1 and sCD14 were measured as described in patients and methods. Each dot corresponds to individual patients with cirrhosis (n=45). Analysis was performed in samples collected from peripheral veins (n=25, ) and from hepatic veins (n=20, )

Article Snippet: For sCD14, human CD14 ELISA Development kit (R&D Systems, Abingdon, UK) was used following manufacturer’s instructions.

Techniques:

Journal: Annals of Gastroenterology : Quarterly Publication of the Hellenic Society of Gastroenterology

Article Title: Systemic levels of human β-defensin 1 are elevated in patients with cirrhosis

doi:

Figure Lengend Snippet: Soluble CD14 (sCD14) and lipopolysaccharide binding protein (LBP) strongly correlate in serum of patients with cirrhosis. Concentrations of sCD14 were measured as described in Patients and Methods. Concentrations of LBP were measured by a commercially available ELISA, according to manufacturer’s instructions. Each dot corresponds to individual patients with cirrhosis (n=36). Analysis was performed in samples collected from peripheral veins

Article Snippet: For sCD14, human CD14 ELISA Development kit (R&D Systems, Abingdon, UK) was used following manufacturer’s instructions.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Development of a newly immunoassay specific for mouse presepsin (sCD14-ST)

doi: 10.1038/s41598-022-22096-1

Figure Lengend Snippet: Specificity of the established polyclonal antibodies (N-pep2 and C-pep8) for mouse presepsin as assessed by immobilized antigens. ( A ) Sequential diluted polyclonal antibodies (culture supernatant) were incubated with immobilized recombinant mouse presepsin-Fc (sCD14-ST-Fc) and binding antibodies were detected by peroxidase-labeled anti-rabbit immunoglobulins antibody (DAKO, P0448). Binding activity is shown by the absorbance of 450–630 nm. ( B ) Diluted purified polyclonal antibodies were incubated with immobilized recombinant mouse CD14-His and binding antibodies were detected by peroxidase-labeled anti-rabbit immunoglobulins antibody (DAKO, P0448). Binding activity is shown by the absorbance of 492 nm.

Article Snippet: Twenty blood samples were centrifuged at 2000 g for 15 min at 4 °C, and plasma fractions were collected and used to determine the levels of presepsin and sCD14 using the developed mouse presepsin kit and the commercially available Quantikine CD14 ELISA kit (R&D systems, Minneapolis, MN, USA).

Techniques: Incubation, Recombinant, Binding Assay, Labeling, Activity Assay, Purification

Journal: Scientific Reports

Article Title: Development of a newly immunoassay specific for mouse presepsin (sCD14-ST)

doi: 10.1038/s41598-022-22096-1

Figure Lengend Snippet: Representative mouse presepsin standard curve with dynamic range 4.7–300 pg/mL of presepsin-Fc protein of the developed ELISA. Each point representative of duplicate measurements.

Article Snippet: Twenty blood samples were centrifuged at 2000 g for 15 min at 4 °C, and plasma fractions were collected and used to determine the levels of presepsin and sCD14 using the developed mouse presepsin kit and the commercially available Quantikine CD14 ELISA kit (R&D systems, Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Development of a newly immunoassay specific for mouse presepsin (sCD14-ST)

doi: 10.1038/s41598-022-22096-1

Figure Lengend Snippet: Linearity of the developed ELISA for mouse presepsin in serial diluted mouse serum. The three individual serum samples were serially diluted as indicated for determination by the developed ELISA. Each point representative the mean value of duplicate measurements.

Article Snippet: Twenty blood samples were centrifuged at 2000 g for 15 min at 4 °C, and plasma fractions were collected and used to determine the levels of presepsin and sCD14 using the developed mouse presepsin kit and the commercially available Quantikine CD14 ELISA kit (R&D systems, Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Development of a newly immunoassay specific for mouse presepsin (sCD14-ST)

doi: 10.1038/s41598-022-22096-1

Figure Lengend Snippet: Reactivity with recombinant mouse CD14-His and mouse presepsin prepared from mouse CD14-His by incubation with cathepsin D. Two antigens were assessed using the developed ELISA. Absorbance is shown by the mean value of duplicate measurements.

Article Snippet: Twenty blood samples were centrifuged at 2000 g for 15 min at 4 °C, and plasma fractions were collected and used to determine the levels of presepsin and sCD14 using the developed mouse presepsin kit and the commercially available Quantikine CD14 ELISA kit (R&D systems, Minneapolis, MN, USA).

Techniques: Recombinant, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Development of a newly immunoassay specific for mouse presepsin (sCD14-ST)

doi: 10.1038/s41598-022-22096-1

Figure Lengend Snippet: Structure of mouse presepsin and its amino acid sequences in CD14. ( A ) Presepsin standard is a 68-residue-long single polypeptide having two intramolecular disulfide bonds. CD14 estimated to processed by proteolytic cleavage at the cathepsin D cleavage site (64R–65E) to generate presepsin (sCD14-ST) in mice. ( B ) Amino acid sequences of immunized peptide for generate anti-mouse presepsin antibodies.

Article Snippet: Twenty blood samples were centrifuged at 2000 g for 15 min at 4 °C, and plasma fractions were collected and used to determine the levels of presepsin and sCD14 using the developed mouse presepsin kit and the commercially available Quantikine CD14 ELISA kit (R&D systems, Minneapolis, MN, USA).

Techniques: Residue

Journal:

Article Title: Differential Inhibition of TRAIL-Mediated DR5-DISC Formation by Decoy Receptors 1 and 2

doi: 10.1128/MCB.00520-06

Figure Lengend Snippet: DcR1 inhibits TRAIL-DISC formation in lipid rafts. (A) Jurkat and (B) HeLa cell populations were left untreated or stimulated for 1 h with 100 ng/ml of His-TRAIL, lysed in Triton X-100, and subjected to density gradient fractionation. Fractions corresponding to lipid rafts (R) and soluble proteins (S) were immunoblotted for DR5, DcR1, DcR2, and flotillin-1 as indicated. Data are representative of at least three independent experiments. Fractions were analyzed for cholesterol content in Jurkat (C) and (D) HeLa cells by use of the Molecular Probes Amplex Red cholesterol assay kit. (E) TRAIL DISC formation in Jurkat cell populations was analyzed after Flag-TRAIL stimulation and immunoprecipitation (IP) in the presence of the M2 anti-Flag antibody as in Fig. ​Fig.2,2, for the indicated time, from cell lysates in (E) NP-40- or (F) Brij78-containing lysis buffer. (G) Analysis of TRAIL protein content from density sucrose gradient fractions obtained from Jurkat cells treated or not treated with His-TRAIL, as described for panel A, was performed by use of an ELISA kit obtained from Diaclone (see Materials and Methods). (H) Freshly isolated monocytes were analyzed by fluorescence-activated cell sorter staining for CD14 and TRAIL receptor expression as indicated. (I) DcR1 protein content from freshly isolated monocytes was analyzed by use of a DcR1 ELISA kit from Diaclone after density sucrose gradient fractionation. Lipid rafts (R) and soluble protein fractions (S) are indicated. (J) Monocytes were stimulated with His-TRAIL as described for panel A, and TRAIL was measured by ELISA, as described above, in lipid rafts (R) and soluble protein fractions (S) as indicated.

Article Snippet: Fractions were analyzed for cholesterol content in Jurkat (C) and (D) HeLa cells by use of the Molecular Probes Amplex Red cholesterol assay kit. (E) TRAIL DISC formation in Jurkat cell populations was analyzed after Flag-TRAIL stimulation and immunoprecipitation (IP) in the presence of the M2 anti-Flag antibody as in Fig. , for the indicated time, from cell lysates in (E) NP-40- or (F) Brij78-containing lysis buffer. (G) Analysis of TRAIL protein content from density sucrose gradient fractions obtained from Jurkat cells treated or not treated with His-TRAIL, as described for panel A, was performed by use of an ELISA kit obtained from Diaclone (see Materials and Methods). (H) Freshly isolated monocytes were analyzed by fluorescence-activated cell sorter staining for CD14 and TRAIL receptor expression as indicated. (I) DcR1 protein content from freshly isolated monocytes was analyzed by use of a DcR1 ELISA kit from Diaclone after density sucrose gradient fractionation.

Techniques: Fractionation, Amplex Red Cholesterol Assay, Immunoprecipitation, Lysis, Enzyme-linked Immunosorbent Assay, Isolation, Fluorescence, Staining, Expressing