Journal: JCI Insight

Article Title: Identifying a biological signature of prenatal maternal stress

doi: 10.1172/jci.insight.143007

Figure Lengend Snippet: ( A ) Heatmap of normalized biomarker means across scoring groups and time points. Individual boxes are represented on a scale from red to blue. Darker red boxes represent markers whose concentrations were found to be at least 1 standard deviation lower than the mean concentration across stress group and time point. Darker blue boxes represent markers whose concentrations were found to be at least 1 standard deviation higher than the mean concentration across stress group and time point. Lower z scores are evident for lipopolysaccharide binding protein (LBP), soluble CD14 (sCD14), and TNF-α in the low-scoring group at 15 and 20 weeks; and for IL-8 in the high-scoring group at 15 and 20 weeks. Significant differences between PSS-scoring groups are illustrated for ( B ) LBP, ( C ) sCD14, ( D ) IL-8, and ( E ) TNF-α (2-way ANOVA with Tukey’s honestly significant differences test, HSD). * P < 0.05.

Article Snippet: Levels of circulating IFABP and sCD14 (Quantikine Immunoassays, R&D Systems, Bio-Techne), LBP (Hycult Biotech), and anti-endotoxin core antibodies (EndoCAb IgA IgG IgM, Hycult Biotech) were determined using commercially available quantitative ELISAs.

Techniques: Biomarker Assay, Standard Deviation, Concentration Assay, Binding Assay

Journal: JCI Insight

Article Title: Identifying a biological signature of prenatal maternal stress

doi: 10.1172/jci.insight.143007

Figure Lengend Snippet: ( A ) Heatmap of normalized biomarker means across scoring groups and time points. Individual boxes are represented on a scale from red to blue. Darker red boxes represent markers whose concentrations were found to be greater than 1 standard deviation lower than the mean concentration across stress group and time point. Darker blue boxes represent markers whose concentrations were found to be greater than 1 standard deviation higher than the mean concentration across stress group and time point. Lower z scores are evident for intestinal fatty acid binding protein (IFABP) in the low-scoring group at 20 weeks. Higher z scores are evident for IL-6 in the low-scoring group at 15 weeks; for IFABP, anti-endotoxin IgG, IL-8, and tryptophan/kynurenine ratio (Trp/Kyn ratio) in the high-scoring group at 15 weeks; and for LBP, C-reactive protein (CRP), and IFN-γ–induced protein 10 (IP-10) in the high-scoring group at 20 weeks. Significant differences between STAI-scoring groups are illustrated for ( B ) LBP, ( C ) sCD14, and ( D ) CRP (2-way ANOVA with Tukey’s HSD). * P < 0.05. MCP-1, monocyte chemoattractant protein-1; MIF, macrophage migration inhibitory factor; SDF-1α, stromal cell-derived factor 1α.

Article Snippet: Levels of circulating IFABP and sCD14 (Quantikine Immunoassays, R&D Systems, Bio-Techne), LBP (Hycult Biotech), and anti-endotoxin core antibodies (EndoCAb IgA IgG IgM, Hycult Biotech) were determined using commercially available quantitative ELISAs.

Techniques: Biomarker Assay, Standard Deviation, Concentration Assay, Binding Assay, Migration, Derivative Assay

Journal: JCI Insight

Article Title: Identifying a biological signature of prenatal maternal stress

doi: 10.1172/jci.insight.143007

Figure Lengend Snippet: ( A ) Heatmap of normalized biomarker means across scoring groups and time points. Individual boxes are represented on a scale from red to blue. Darker red boxes represent markers whose concentrations were found to be greater than 1 standard deviation lower than the mean concentration across stress group and time point. Darker blue boxes represent markers whose concentrations were found to be greater than 1 standard deviation higher than the mean concentration across stress group and time point. Lower z scores are evident for LBP, sCD14, and TNF-α in the low-scoring group at 15 and 20 weeks and for IL-8 and SDF-1α in the high-scoring group at 20 weeks. Higher z scores are evident for IFABP and Trp/Kyn ratio in the high-scoring group at 15 weeks and for CRP in the high-scoring group at 15 and 20 weeks. Significant differences between EPDS-scoring groups are illustrated for ( B ) LBP, ( C ) sCD14, ( D ) CRP, ( E ) TNF-α, and ( F ) IL-8 (2-way ANOVA with Tukey’s HSD). * P < 0.05.

Article Snippet: Levels of circulating IFABP and sCD14 (Quantikine Immunoassays, R&D Systems, Bio-Techne), LBP (Hycult Biotech), and anti-endotoxin core antibodies (EndoCAb IgA IgG IgM, Hycult Biotech) were determined using commercially available quantitative ELISAs.

Techniques: Biomarker Assay, Standard Deviation, Concentration Assay

Journal: JPEN. Journal of parenteral and enteral nutrition

Article Title: Faecalibacterium prausnitzii and a Prebiotic Protect Intestinal Health in a Mouse Model of Antibiotic and Clostridium difficile Exposure

doi: 10.1002/jpen.1053

Figure Lengend Snippet: Effects of Clostridium difficile (CD) on tight junction protein and an anion exchanger expression in proximal colon. Mice were treated as described in Figures 1 and ​and2.2. Proximal colon was collected and embedded in optimal cutting temperature medium (OCT) for histology on days 1 and 5 after CD exposure. (A) Occludin (red), zona occludin-1 (ZO-1; green), (B) claudin-3 (green), and (C) Na+/H+ exchanger isoform 3 (NHE3; green) were visualized by immunohistochemistry in sections of proximal colon frozen in OCT. A selected area was cropped and enlarged. All images were acquired using a 40 × objective. Images are representative of at least replicate images captured per mouse in 3–6 mice per treatment group. FP, Faecalibacterium prausnitzii; PS, potato starch.

Article Snippet: Antibodies were purchased from the following sources: anti–Na + /H + exchanger isoform 3 (NHE 3 ), anti–zona occludin-1 (ZO-1), anti–sodium-coupled monocarboxylate transporter 1 (SLC5A8), and anti–frizzled class receptor 7 (FZD7) from Abcam (Cambridge, MA); anti-occludin from Hycult Biotechnologies (Plymouth Meeting, PA); anti-claudin-3, Alexa Fluor 488, and 568 IgG from Invitrogen (Camarillo, CA).

Techniques: Expressing, Immunohistochemistry

Journal: World Journal of Gastroenterology

Article Title: Gut barrier failure biomarkers are associated with poor disease outcome in patients with primary sclerosing cholangitis

doi: 10.3748/wjg.v23.i29.5412

Figure Lengend Snippet: Serum level of bacterial translocation and enterocyte damage markers according to the different serologic antibody statuses

Article Snippet: Presence of IgA or IgG type antibodies against F-actin (AAA) and gliadin (AGA) (QUANTA Lite ® ; INOVA Diagnostics, San Diego, CA) and level of intestinal fatty acid-binding protein (I-FABP) (Hycult Biotechnology, Uden, Netherlands) were determined as the serologic markers of gut barrier function.

Techniques: Translocation Assay

Journal: Alcoholism, clinical and experimental research

Article Title: Specifically-sized hyaluronan (35 kDa) prevents ethanol-induced disruption of epithelial tight junctions through a Layilin-dependent mechanism in Caco-2 cells

doi: 10.1111/acer.14140

Figure Lengend Snippet: (A/B/C) Caco-2 monolayers were differentiated for 21 days and then treated or not with 100 μg/ml HA35 for 24 hours, followed by challenge with or without 40 mM ethanol for 3 hours. (A) Cells were then fixed in methanol and immunoreactive ZO-1 (green) and occludin (red) visualized by confocal microscopy. Images acquired at 63X with a 3X digital magnification and are representative of at least two images per slide from three independent experiments. (B) Occludin and ZO-1 expression was assessed by immunoblot using ß-actin as a loading control. Semi-quantification of protein expression was performed using Carestream Imaging Software. (C) FITC-dextran (4Kd) was added to the upper chamber of the transwells and leakage into the lower chamber assessed at 1 and 2h. (D) Differentiated Caco-2 monolayers were treated with increasing concentrations of HA35 for 24 h and then challenged with 500 mM ethanol. TEER was measured immediately before the addition of ethanol and 3 h later. Values are expressed as means ±SEM, (A) n=3, (B) n=4–6 and (C) n=12 (values with different superscripts are significantly different within a time point, p<0.05) and (D) n=4 (*p < 0.05 compared to cells not treated with HA35)

Article Snippet: Primary antibodies were purchased from the following sources: rabbit anti-ZO-1 (Life Technologies, 61–7300), guinea pig anti-occludin (Hycult HP9047), rabbit anti-ß-actin (Cell Signaling, 4967), rabbit anti-Layilin (Bioss, 1842R).

Techniques: Confocal Microscopy, Expressing, Western Blot, Imaging, Software

Journal: Alcoholism, clinical and experimental research

Article Title: Specifically-sized hyaluronan (35 kDa) prevents ethanol-induced disruption of epithelial tight junctions through a Layilin-dependent mechanism in Caco-2 cells

doi: 10.1111/acer.14140

Figure Lengend Snippet: Caco-2 monolayers were differentiated on collagen-coated transwell filters and then transfected with scrambled siRNA or siRNA targeting Layilin. (A) Immunofluorescent staining for Layilin 24 h following transfection with scrambled siRNA or siRNA targeting Layilin. Images are representative of at least two images per slide from three independent transfections. (B) 24 h after transfection, cells were treated or not with 100μg/ml HA35 for 24 hours and then challenged with or without 40mM ethanol for 3 hours. Immunoreactive ZO-1 (green) and occludin (red) was visualized by confocal microscopy in cells transfected with scrambled siRNA or siRNA targeting Layilin. Images acquired at 63X with a 3X digital magnification and are representative of at least two images per slide from four independent experiments. (C) Cells transfected with scrambled siRNA or siRNA targeting Layilin were treated with 1 mg/ml HA35 for 24 h and then challenged with 500 mM ethanol. TEER was measured immediately after the addition of ethanol and 3 h later. Values are expressed as means ± SEM. Values are expressed as means ±SEM, (A) n=3, (B) n=6 and (C) n=4–6, *p < 0.05 compared to cells not treated with ethanol within a transfection group.

Article Snippet: Primary antibodies were purchased from the following sources: rabbit anti-ZO-1 (Life Technologies, 61–7300), guinea pig anti-occludin (Hycult HP9047), rabbit anti-ß-actin (Cell Signaling, 4967), rabbit anti-Layilin (Bioss, 1842R).

Techniques: Transfection, Staining, Confocal Microscopy