Journal: Frontiers in Microbiology

Article Title: Presence of Segmented Filamentous Bacteria in Human Children and Its Potential Role in the Modulation of Human Gut Immunity

doi: 10.3389/fmicb.2018.01403

Figure Lengend Snippet: (A) Overall relationship between the level of sIgA in luminal fluids from the SFB-positive and SFB-negative patients. (B) Relationship between the level of sIgA and the ages of children. A total number of 47 samples (23 SFB-positive and 24 SFB-negative) were used. 500 μl of luminal fluids from each patient were used for the measurement of sIgA as described in the Section “Materials and Methods.” The supernatants were assayed for total secretory IgA using a human IgA ELISA Detection Kit as described in the section “Materials and Methods.” The sIgA was expressed as μg/ml of fecal materials. P -values were calculated using Student’s t -test.

Article Snippet: Samples were incubated at room temperature for 2 h. Total sIgA was quantified by ELISA using a human secretory IgA ELISA detection Kit (Elabscience, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Dose-dependent cytopathic effect of rotavirus on bovine intestinal epithelial cells (CIEB), pig intestinal epithelial cells (CLAB), and human intestinal epithelial cells (H4). The graphs for cytopathic effect of rotavirus on the different cells are shown in terms of mean absorbance at each concentration ± standard deviation at 595 nm.

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Concentration Assay, Standard Deviation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Survival of bovine intestinal epithelial cells (CIEB), pig intestinal epithelial cells (CLAB), and human intestinal epithelial cells (H4) from cytopathic effects of rotavirus in the presence of T. esculentum extracts. Shown above are bars of % survival of epithelial cells {% change in cell viability after coincubation of rotavirus with marama extracts compared to rotavirus only wells at OD 595 nm ± % standard deviation}, respective levels of significance in terms of T values (*). T values were determined using Statsoft Statistica software ( P < .05). Tests were done in triplicate wells. * T value between 0 and 5 (low level significance), ** T value greater than 5, but less than 10 (middle level significance), and *** T value greater than 10 (high level significance). MCE: T. esculentum bean cotyledon ethanolic extract, MSCE: T. esculentum bean husk ethanolic extract, MTW: T. esculentum tuber water extract, MSCW: T. esculentum bean husk water extract, MCW: T. esculentum bean cotyledon water extract, and Combivir: (lamivudine and zidovudine).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Standard Deviation, Software

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: T. esculentum extracts show direct inhibition of rotavirus infectivity on bovine intestinal epithelial cells (CIEB), pig intestinal epithelial cells (CLAB), and human intestinal epithelial cells (H4). Shown above are bars of % survival of epithelial cells {% change in cell viability after pre-exposure of rotavirus to T. esculentum extracts compared to rotavirus only wells at OD 595 nm ± % standard deviation}, respective levels of significance in terms of T values (*). T values were determined using Statsoft Statistica software ( P < .05). Tests were done in triplicate wells. * T value between 0 and 5 (low level significance), ** T value greater than 5, but less than 10 (middle level significance), and *** T value greater than 10 (high level significance). MCE: T. esculentum bean cotyledon ethanolic extract, MSCE: T. esculentum bean husk ethanolic extract, MTW: T. esculentum tuber water extract, MSCW: T. esculentum bean husk water extract, MCW: T. esculentum bean cotyledon water extract, and AZT: Combivir (lamivudine and zidovudine).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Inhibition, Infection, Standard Deviation, Software

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Effect of coincubation of rotavirus with T. esculentum extracts on monolayer polarity (TER) (Ω/cm 2 ) of H4 cells over time. Shown graphs of change in epithelial cell monolayer polarity exposed to extracts or AZT and rotavirus (treatment wells), extracts or AZT alone (control), or rotavirus alone (negative control) over time compared to respective controls. Tests were done in triplicate wells (12-well Transwell plates). MCE: T. esculentum bean cotyledon ethanolic extract (1 mg/mL), MSCE: T. esculentum bean husk ethanolic extract (0.1 mg/mL), MTW: T. esculentum tuber water extract (0.1 mg/mL), MSCW: T. esculentum bean husk water extract (0.01 mg/mL), MCW: T. esculentum bean cotyledon water extract (2 mg/mL), and AZT: Combivir (lamivudine and zidovudine) (0.75 mg/mL).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Control, Negative Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Tylosema esculentum (Marama) Tuber and Bean Extracts Are Strong Antiviral Agents against Rotavirus Infection

doi: 10.1155/2011/284795

Figure Lengend Snippet: Effect of T. esculentum extracts on release of nitric oxide (NO) (% release compared to control) in human small intestine epithelial cells (H4) and pig small intestine epithelial cells (CLAB). MCE: T. esculentum bean cotyledon ethanolic extract (1 mg/mL), MSCE: T. esculentum bean husk ethanolic extract (0.1 mg/mL), MTW: T. esculentum tuber water extract (0.1 mg/mL), MSCW: T. esculentum bean husk water extract (0.01 mg/mL), and MCW: T. esculentum bean cotyledon water extract (2 mg/mL).

Article Snippet: NO 2 release was significant from human intestinal epithelial cells (H4) exposed to all extracts used {seed coat water and ethanolic extracts (MSCW and MSCE, resp.), cotyledon water and ethanolic extracts (MCW and MCE, resp.), and tuber water extract (MTW)}, while only MCE and MTW significantly increased NO in CLAB cells ( ).

Techniques: Control

Journal: Nature Communications

Article Title: Structural plasticity of KIR2DL2 and KIR2DL3 enables altered docking geometries atop HLA-C

doi: 10.1038/s41467-021-22359-x

Figure Lengend Snippet: a Overall structure of KIR2DL3 in complex with HLA-C*07:02-RL9. The complex is represented in cartoon format with KIR2DL2 coloured blue, HLA-C*07:02 coloured grey, β2 m coloured orange and the peptide coloured cyan. b Comparison of KIR2DL2 (pink) and 2DL3 (blue) in complex with HLA-C*07:02-RL9. The relative difference in hinge and twist angles of the KIR is shown. c Footprint of KIR2DL3 on HLA-C*07:02-RL9 coloured by domain binding. D1 contacts coloured green, D1-D2 interdomain loop contacts coloured red and D2 contacts coloured orange. d Comparison of KIR2DL2 (pink) and KIR2DL3 (blue) D1 contacts to the α1-helix of HLA-C*07:02-RL9 (KIR2DL2 contacts coloured grey, KIR2DL3 contacts coloured cyan). The ~3 Å shifts of the D1 C-C′ and E-F loops are highlighted. e Mapping of sequence differences to the structures of KIR2DL2/3. Differences highlighted as blue spheres.

Article Snippet: Single-cell suspension was incubated in staining antibody cocktail containing a combination of following antibodies: CD56 BV421 (clone NCAM16.2; BD Biosciences; 1:50 dilution), CD56 APC or PE (clone B159; BD Biosciences; 1:50 dilution), KIR3DL1 APC (clone DX9; BD Biosciences 1:50 dilution), CD3 APCCy7 (clone SK7; BD Biosciences; 1:100 dilution), KIR2DL2/L3/S2 PECy5.5 (clone GL183; Beckman Coulter; 1:50-1:200 dilution), KIR2DL2/L3/S2 BUV737 (clone CH-L; BD Biosciences; 1:50 dilution), KIR2DL1/S1 PECy7 or APC (clone EB6; Beckman Coulter; 1:50 dilution), KIR2DL3 FITC (clone REA147; Miltenyi Biotech; 1:50 dilution), fixable viability dye eFluor780 (Life Research; 1:500 dilution).

Techniques: Comparison, Binding Assay, Sequencing

Journal: Nature Communications

Article Title: Structural plasticity of KIR2DL2 and KIR2DL3 enables altered docking geometries atop HLA-C

doi: 10.1038/s41467-021-22359-x

Figure Lengend Snippet: 293T cells were transfected with plasmids encoding either wild-type or mutant FLAG-tagged KIR2DL2 and KIR2DL3 and then stained with a FLAG-specific mAb or HLA-C*03:04-GL9 and HLA-C*07:02-RL9 tetramers. Representative dot plots showing staining to cells expressing wild-type KIR2DL2 and KIR2LD3 ( a ). Relative binding of wild-type (WT) KIR2DL2 and KIR2DL3 transfected 293T cells and their respective alanine mutants to phycoerythrin-conjugated tetramers of HLA-C*03:04-GL9 ( b ) and HLA-C*07:02-RL9 ( c ). Transfected cells were stained for FLAG as a marker of KIR2DL expression and separately stained with HLA tetramers. Percentage tetramer+ was normalised to percentage FLAG + to account for differing KIR2DL transfection efficiency between mutants. The data represent a minimum of n = 3 independent experiments; bar line = mean, error bars = standard error of the mean (SEM); individual data points = circles (2DL2) or squares (2DL3). For each independent replicate, relative tetramer staining was normalised to staining of WT KIR2DL. Statistically significant differences in ligand recognition between corresponding mutants of KIR2DL2 and KIR2DL3 as measured by Two-way ANOVA Šidák’s multiple comparison test are highlighted. 2DL2 ( n = 5) vs. 2DL3 ( n = 3) binding to C*03:04-GL9 K44A: * P = 0.0127; 2DL2 ( n = 5) vs. 2DL3 ( n = 3) binding to C*03:04-GL9 S133A: *** P = 0.0005; 2DL2 ( n = 5) vs. 2DL3 ( n = 3) binding to C*07-02-GL9 D135A: *** P = 0.0007.

Article Snippet: Single-cell suspension was incubated in staining antibody cocktail containing a combination of following antibodies: CD56 BV421 (clone NCAM16.2; BD Biosciences; 1:50 dilution), CD56 APC or PE (clone B159; BD Biosciences; 1:50 dilution), KIR3DL1 APC (clone DX9; BD Biosciences 1:50 dilution), CD3 APCCy7 (clone SK7; BD Biosciences; 1:100 dilution), KIR2DL2/L3/S2 PECy5.5 (clone GL183; Beckman Coulter; 1:50-1:200 dilution), KIR2DL2/L3/S2 BUV737 (clone CH-L; BD Biosciences; 1:50 dilution), KIR2DL1/S1 PECy7 or APC (clone EB6; Beckman Coulter; 1:50 dilution), KIR2DL3 FITC (clone REA147; Miltenyi Biotech; 1:50 dilution), fixable viability dye eFluor780 (Life Research; 1:500 dilution).

Techniques: Transfection, Mutagenesis, Staining, Expressing, Binding Assay, Marker, Comparison

Journal: Nature Communications

Article Title: Structural plasticity of KIR2DL2 and KIR2DL3 enables altered docking geometries atop HLA-C

doi: 10.1038/s41467-021-22359-x

Figure Lengend Snippet: KIR2DL2 and KIR2DL3 binding to HLA-C*07:02-RL9-P7/P8 mutant peptide panel.

Article Snippet: Single-cell suspension was incubated in staining antibody cocktail containing a combination of following antibodies: CD56 BV421 (clone NCAM16.2; BD Biosciences; 1:50 dilution), CD56 APC or PE (clone B159; BD Biosciences; 1:50 dilution), KIR3DL1 APC (clone DX9; BD Biosciences 1:50 dilution), CD3 APCCy7 (clone SK7; BD Biosciences; 1:100 dilution), KIR2DL2/L3/S2 PECy5.5 (clone GL183; Beckman Coulter; 1:50-1:200 dilution), KIR2DL2/L3/S2 BUV737 (clone CH-L; BD Biosciences; 1:50 dilution), KIR2DL1/S1 PECy7 or APC (clone EB6; Beckman Coulter; 1:50 dilution), KIR2DL3 FITC (clone REA147; Miltenyi Biotech; 1:50 dilution), fixable viability dye eFluor780 (Life Research; 1:500 dilution).

Techniques: Binding Assay, Mutagenesis

Journal: Nature Communications

Article Title: Structural plasticity of KIR2DL2 and KIR2DL3 enables altered docking geometries atop HLA-C

doi: 10.1038/s41467-021-22359-x

Figure Lengend Snippet: Enriched NK cells from healthy donors were cultured with target cells and the degranulation response of NK cells expressing KIR2D assessed by flow cytometry. a The expression of HLA-C on both 721.221 and Raji, Ramos and Namalwa lymphoma cell lines following staining with DT9 antibody (shaded grey area) is shown, with secondary antibody alone shown as unfilled histogram. b Gating strategy for KIR2DL1/S1, KIR2DL2/S2 and KIR2DL3 NK cells. c Representative flow cytometry analysis of CD107a expression on NK cell subsets from one donor in the presence of 721.221, Raji, Ramos and Namalwa cells is shown (gated on KIR3DL1 − NK cells). d Pooled data showing CD107a+ cells from six donors with six KIR2DL1/S1, and KIR2DL3 populations and four KIR2DL2/S2 populations is shown. Each symbol represents an individual population of cells and bars depict mean ± SEM values. CD107a response is normalised to 721.221 response from the same experiment. e Representative CD107a expression on NK cell subsets from one donor in the presence of either 721.221 cells or 721.221 cells transfected HLA-C*06:02 or -C*07:02. f The degranulation response of KIR2D + NK cells (GL183+ gated on a set GL183 MFI) to untransfected and HLA-C transfected 721.221 cells. Data are pooled from five donors. Each symbol represents data from an individual donor and bars depict mean ± SEM values. CD107a response is normalised to 721.221 response. g The expression of different HLA-C on target cells stained with W6/32 antibody (shaded grey area) is shown with 721.221 stain shown as black line. Statistical significance between KIR3DL2+ and KIR2DL3+ subsets (excluding KIR2DL1+) was tested using Mann–Whitney test for all conditions; no symbol = p > 0.05; * p < 0.05; ** p < 0.01. Each experiment has been performed twice with data from both experiments pooled for the figure.

Article Snippet: Single-cell suspension was incubated in staining antibody cocktail containing a combination of following antibodies: CD56 BV421 (clone NCAM16.2; BD Biosciences; 1:50 dilution), CD56 APC or PE (clone B159; BD Biosciences; 1:50 dilution), KIR3DL1 APC (clone DX9; BD Biosciences 1:50 dilution), CD3 APCCy7 (clone SK7; BD Biosciences; 1:100 dilution), KIR2DL2/L3/S2 PECy5.5 (clone GL183; Beckman Coulter; 1:50-1:200 dilution), KIR2DL2/L3/S2 BUV737 (clone CH-L; BD Biosciences; 1:50 dilution), KIR2DL1/S1 PECy7 or APC (clone EB6; Beckman Coulter; 1:50 dilution), KIR2DL3 FITC (clone REA147; Miltenyi Biotech; 1:50 dilution), fixable viability dye eFluor780 (Life Research; 1:500 dilution).

Techniques: Cell Culture, Expressing, Flow Cytometry, Staining, Transfection, MANN-WHITNEY

Journal: Frontiers in Medicine

Article Title: Predicting cardiometabolic disease in medical students using FibroScan and 30-year Framingham risk scores

doi: 10.3389/fmed.2024.1431935

Figure Lengend Snippet: Comparison of participants with and without elevated liver stiffness.

Article Snippet: To measure human immunoglobulin A2 and human PIGR levels, we used a commercially available enzyme-linked immunosorbent assay kit from Elabscience Biotechnology Inc., United States.

Techniques: Comparison, Control, Clinical Proteomics