Journal: Journal of biomedical materials research. Part A

Article Title: Artificial Lymphatic Drainage Systems for Vascularized Microfluidic Scaffolds

doi: 10.1002/jbm.a.34524

Figure Lengend Snippet: Endothelial delamination in the absence of drainage. (A) Relevant dimensions of the vascularized, undrained fibrin scaffold. Microvessels were constructed by growing blood microvessel-derived endothelial cells in microfluidic channels (120 μm diameter) inside fibrin gels. The length of microvessels was 8.1 ± 0.3 mm, and the thickness of the scaffold was 1.1 mm. (B) Phase-contrast images of microvessels that were perfused for 3 days in fibrin scaffolds of the indicated concentrations. (C) Brightfield images of microvessels that were perfused for 9 days; endothelial delamination is indicated. (D–E) Method for generating delamination frequency maps. (D) Black and white bars were used to highlight stable and delaminated regions, respectively, producing binary delamination maps along the entire length (as shown in (E)) on each side of the vessel; as such, two maps were obtained per vessel. (E) These binary maps were then stacked and their intensity averaged, to generate delamination frequency maps. (F) Frequency maps of delamination (6 mg/mL, 2n = 30; 8 mg/mL, 2n = 22; 10 mg/mL, 2n = 22; 15 mg/mL, 2n = 24; 30 mg/mL, 2n = 34). Brighter regions indicate higher delamination frequency. (G) Delaminated length expressed as a fraction of the total length. (H) Hydraulic conductivities of fibrin gels. (I) Flow rates of microvessels in 30 mg/mL scaffolds in the undrained and T-junction drainage conditions (δ = 0.5 mm and 6.5 mm).

Article Snippet: Cell culture We cultured endothelial cells derived from human dermal blood microvessels (lot 0040804.2 from PromoCell, and lot 5F1293 from Lonza) at 37°C in 5% CO 2 .

Techniques: Construct, Derivative Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: RSV induces TSLP expression in primary AECs. A, NHBECs were exposed to UV-irradiated RSV or RSV with indicated virus titers for 12 hours, and TSLP mRNA was measured by real-time quantitative PCR for virus titer. B and C, NHBECs were exposed to UV-irradiated RSV or RSV (MOI, 1) for indicated time course. Cell or culture supernatant fluids were harvested, and TSLP mRNA was measured by real-time quantitative PCR (Fig 1, B). TSLP protein levels were measured by ELISA (Fig 1, C). D and E, NHBECs were exposed to RSV (MOI, 1) without or with anti-TNFα antibodies (10, 50, or 100 ng/mL) (Fig 1, E), anti–IL-1R (1, 10, or 50 µg/mL) (Fig 1, E) for 24 hours, and culture supernatant fluids were harvested. TSLP protein levels were analyzed by ELISA. Data represent means ± SDs of 3 independent measurements. Similar results were obtained for 5 independent experiments (Fig 1, A–E). *P ≤ .05.

Article Snippet: Cells and virus Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Expressing, Irradiation, Virus, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: RIG-I and IPS-1 activation mediates Paramyxovirus-induced TSLP expression. A and B, A549 cells were transfected with control plasmid or luciferase constructs containing the human TSLP promoter alone or in combination with expression vectors for DN-RIG-I (Fig 2, A) or IPS-1 (Fig 2, B) followed by infection with RSV (MOI, 1). C, Normal littermate control (NLC) or RIG-I KO MEFs were infected with or without RSV, and TSLP protein expression was measured by ELISA. n.d. indicates not detected. D, NHBECs were transfected with control or RIG-I siRNA, followed by RSV infection, and TSLP mRNA was assessed at 8 hours after infection. Data represent means ± SDs of 3 independent measurements. Similar results were obtained for 5 independent experiments (Fig 2, A–D). *P ≤ .05.

Article Snippet: Cells and virus Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Activation Assay, Expressing, Transfection, Control, Plasmid Preparation, Luciferase, Construct, Infection, Enzyme-linked Immunosorbent Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: Human asthmatic epithelium produces greater levels of TSLP in response to RSV infection. ALI cultures were generated from primary BECs isolated from healthy or asthmatic children via bronchial brushing. Data show TSLP protein levels, measured by ELISA, in basolateral culture supernatant fluids after infection with either RSV Line 19 or RSV A2 or control at an MOI of 0.5 (n = 12 patients for all groups). P values were calculated with an ANOVA with Tukey posttest.

Article Snippet: Cells and virus Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Infection, Generated, Isolation, Enzyme-linked Immunosorbent Assay, Control

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: RSV-mediated expression of RIG-I and TLR3 in ALI cultures from healthy and asthmatic children. RIG-I and TLR3 expression (log2 scale) by BECs from healthy (open plots; n = 9) and asthmatic children (solid plots; n = 12) in response to RSV infection by A2 and Line 19 strains or exposure to control vero cell supernatant fluid (VC). RIG-I expression: ANOVA P < .001 between groups; asthma RSV A2 group 3-fold greater RIG-I expression than asthma VC group (*P < .001); asthma RSV 19 group 4-fold greater RIG-I expression than asthma VC group (*P < .001); no significant change in RIG-I expression by healthy cells with RSV A2 or RSV 19; no significant difference in RIG-I expression between asthmatic and healthy VC groups. TLR3 expression: ANOVA P = .003 between groups; healthy RSV A2 group 1.4-fold greater TLR3 expression than by healthy VC group (**P = .05); healthy RSV 19 group 1.8-fold greater TLR3 expression than by healthy VC group (***P = .03); asthma RSV A2 group 1.5-fold greater TLR3 expression than by asthma VC group (†P = .002); asthma RSV 19 group 1.8-fold greater TLR3 expression than by asthma VC group (††P = .002); no significant difference in TLR3 expression between asthmatic and healthy VC groups. RIG-I and TLR3 expression normalized to GAPDH. Bottom and top of box plots represent 25th and 75th percentiles, respectively; dotted band represents the median and whiskers represent minimums and maximums.

Article Snippet: Cells and virus Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Expressing, Infection, Control

Journal: Clinical Science (London, England : 1979)

Article Title: Down-regulation of STAT3 enhanced chemokine expression and neutrophil recruitment in biliary atresia

doi: 10.1042/CS20201366

Figure Lengend Snippet: ( A ) HE staining showed the cellular infiltrate in CC and BA patient liver samples. The scale bar represents 50 μm. ( B ) Picrosirius Red staining shows fibrosis in CC and BA patient liver samples. ( C ) mRNA expression of STAT3 measured by quantitative PCR. *** P <0.001 ( n =20 for CC and n =28 for BA). ( D ) Protein expression of STAT3 and the activated form p-STAT3 determined by Western blotting. N =4 for both CC and BA; one set of representative results is shown. ( E ) Immunohistochemical staining showed expression of STAT3 and p-STAT3 in BA and CC patient liver samples. The portal areas are shown, and the arrows indicate the positions where BECs can be observed. The solid arrows indicate these features in the CC samples, and the dashed arrows indicate these features in BA. The scale bar represents 50 μm.

Article Snippet: Human intrahepatic BECs were purchased from ScienCell (ScienCell, CA, U.S.A.).

Techniques: Staining, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining

Journal: Clinical Science (London, England : 1979)

Article Title: Down-regulation of STAT3 enhanced chemokine expression and neutrophil recruitment in biliary atresia

doi: 10.1042/CS20201366

Figure Lengend Snippet: ( A ) Human BECs were cultured as described previously. Neutrophils isolated from PBMCs were activated with PMA and added to the BEC cultures for another 24 or 48 h, and then the cultures were photographed. ( B ) Apoptosis in single-BEC suspensions cocultured with activated neutrophils was analysed by flow cytometry. At least three sets of experiments were performed, and one representative graph is shown. ( C ) Apoptosis in BECs co-cultured at 24 and 48 h with activated neutrophils was evaluated. *** P <0.001.

Article Snippet: Human intrahepatic BECs were purchased from ScienCell (ScienCell, CA, U.S.A.).

Techniques: Cell Culture, Isolation, Flow Cytometry

Journal: Clinical Science (London, England : 1979)

Article Title: Down-regulation of STAT3 enhanced chemokine expression and neutrophil recruitment in biliary atresia

doi: 10.1042/CS20201366

Figure Lengend Snippet: ( A ) Human BECs were cultured in epithelial cell medium for 48 h, and then different concentrations of Stattic were added to the cultures and incubated for another 24 h before cell morphology was analysed. ( B,C ) Western blotting shows the expression of p-STAT3 and STAT3 in cell cultures treated with different concentrations of Stattic for 12 h. Cont., control group with Stattic; Stattic 5, Stattic at a dose of 5 μM. ( D ) mRNA expression of chemoattractant IL-8 in human BEC cultures was measured by quantitative PCR. The cont.2.5 and cont.5 group correspond to the control groups for the STAT3 inhibitor Stattic with 2.5 and 5 μM, with the same volume of DMSO as solvent of Stattic. *** P <0.001. ( E ) IL-8 secretion into the supernatant of the cell culture was assessed by ELISA, * P <0.05, *** P <0.001.

Article Snippet: Human intrahepatic BECs were purchased from ScienCell (ScienCell, CA, U.S.A.).

Techniques: Cell Culture, Incubation, Western Blot, Expressing, Control, Real-time Polymerase Chain Reaction, Solvent, Enzyme-linked Immunosorbent Assay