Journal: medRxiv

Article Title: A Phase I, Prospective, Randomized, Open-labeled Study to Evaluate the Safety, Tolerability, and Immunogenicity of Booster Dose with MVC-COV1901 or MVC-COV1901(Beta) SARS-CoV-2 Vaccine in Adults

doi: 10.1101/2022.08.29.22279317

Figure Lengend Snippet: SARS-CoV-2 spike-specific memory B cell (MBC) response before and after the booster dose. ( A ) Illustration of SARS-CoV-2 spike-specific MBC frequency, measured by ELISpot. In the ELISpot assay, antigen-coated wells were used to assess antigen-specific MBC, PBS-coated wells were used as negative controls, anti-Ig coated wells were used to assess total IgG MBC. The numbers (in italics) of spots and cultured cells incubated in the ELISpot assay were shown below each image. Each spot represents an antibody-secreting cell. The frequency of antigen-specific IgG MBC was calculated as the percentage of total IgG MBC. Subject 02-023 is an adult who had two prior doses of MVC-COV1901 and received a booster dose of MVC-COV1901 containing a Beta variant spike of 15 mcg. V2, the vaccination day; V4, 14 days after the booster dose. ( B) spike-specific MBC frequency in the peripheral blood was measured in those with two (group A) and three (group B) initial doses of MVC-COV1901, before (V2) and 14 days (V4) after the booster dose, with memory B cell ELISpot assay. Wild type (WT), Beta, and Omicron BA.1 spike-specific IgG, IgM or IgA MBC frequencies were shown in mean ± SEM in the figure. Each symbol represents a sample (subject). ( C ) spike-specific MBC frequency in the subgroups, i.e., booster dose with MVC-COV1901 containing Wuhan wild type spike, booster dose with MVC-COV1901 containing Beta variant spike 15 mcg, and booster dose with MVC-COV1901 containing Beta variant spike 25 mcg, of group A and B. Wild type (WT), Beta, and Omicron BA.1 spike-specific IgG, IgM or IgA MBC frequencies were shown in mean ± SEM in the figure. Each symbol represents a sample (subject). ( D ) Relationship of spike-specific IgG MBC frequency and serological neutralisation titre with wild type and Omicron variant BA.4/BA.5 SARS-CoV-2 pseudovirus among group A and B subjects. Linear regression was used to model the relationship between two variables. pNT, pseudovirus neutralisation titre. Mann-Whitney test was used to compare MBC frequencies at two-time points. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. ns, not significant.

Article Snippet: Thawed and rested PBMCs were dispensed at 1 × 10 5 cells per well for IFN-g ELISpot assay (Human IFN-g ELISpot Kit, MABTECH) or 2 × 10 5 cells per well for IL-4 ELISpot assay (Human IL-4 ELISpot Kit, MABTECH).

Techniques: Enzyme-linked Immunospot, Cell Culture, Incubation, Variant Assay, MANN-WHITNEY

Journal: The FASEB Journal

Article Title: Monosodium urate crystals alter the circadian clock in macrophages leading to loss of NLRP3 inflammasome repression: Implications for timing of the gout flare

doi: 10.1096/fj.202202035r

Figure Lengend Snippet: FIGURE 4 The effect of monosodium urate (MSU) crystals on Caspase 1 activity and IL-1β secretion is time-of-day dependent. (A) Schematic of the experimental design to assess the time-of-day differences in NLRP3 inflammasome activity. Following 18 h serum starvation, cells were media changed to serum-replete media and either treated immediately with MSU crystals (500 μg/mL) or rested for 12 h before media-changing again with/without MSU crystal addition. Caspase 1 activity was measured following 12 h of MSU crystal exposure and normalized for differences in cell number as determined by CyQuant assay with reference to a standard curve. (B) The magnitude of Caspase 1 activity induced by MSU crystal exposure in cells at the 0–12 h compared with 12–24 h timepoints. (C) The magnitude of increase in secreted IL-1β levels induced by MSU crystal exposure in THP-1 macrophages at the 0–12 h compared with 12–24 h timepoints. Data shown are mean ± SEM. Data were analyzed by t-test with p < .05 considered statistically significant.

Article Snippet: Secreted IL- 1β protein levels were measured in the cell culture supernatant using a Duoset Human IL- 1β ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Activity Assay, CyQUANT Assay

Journal: The FASEB Journal

Article Title: Monosodium urate crystals alter the circadian clock in macrophages leading to loss of NLRP3 inflammasome repression: Implications for timing of the gout flare

doi: 10.1096/fj.202202035r

Figure Lengend Snippet: FIGURE 6 BMAL1 represses expression of pro-IL-1β and CASP1 but not NLRP3 in THP-1 macrophages BMAL1 was knocked down or over-expressed in THP-1 macrophages using adenoviral-mediated gene delivery. Following gene transduction, cells were serum starved for 18 h then media changed to serum-replete media and either treated immediately with MSU crystals (500 μg/mL) (0–12 h timepoint) or rested for 12 h before media-changing again with/without MSU crystal addition (12–24 h timepoint). NLRP3 expression measured by RT-qPCR following BMAL1 knockdown at (A) the 0–12 h timepoint and (B) 12–24 h timepoint. NLRP3 expression measured by RT-qPCR following BMAL1 overexpression at (C) the 0–12 h timepoint and (D) 12–24 h timepoint. Pro-IL-1β expression measured by RT-qPCR following BMAL1 knockdown at (E) the 0–12 h timepoint and (F) 12–24 h timepoint. Pro-IL-1β expression measured by RT-qPCR following BMAL1 overexpression at (G) the 0–12 h timepoint and (H) 12–24 h timepoint. CASP1 expression measured by RT-qPCR following BMAL1 knockdown at (I) the 0–12 h timepoint and (J) 12–24 h timepoint. CASP1 expression measured by RT-qPCR following BMAL1 overexpression at (K) the 0–12 h timepoint and (L) 12–24 h timepoint. Data shown are mean ± SEM for 3 experimental replicates. Data were analyzed by one-way ANOVA (post-hoc Tukey) with p < .05 considered statistically significant.

Article Snippet: Secreted IL- 1β protein levels were measured in the cell culture supernatant using a Duoset Human IL- 1β ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Expressing, Transduction, Quantitative RT-PCR, Knockdown, Over Expression

Journal: The FASEB Journal

Article Title: Monosodium urate crystals alter the circadian clock in macrophages leading to loss of NLRP3 inflammasome repression: Implications for timing of the gout flare

doi: 10.1096/fj.202202035r

Figure Lengend Snippet: FIGURE 7 REV-ERBα represses NLRP3 expression and inflammasome activity in THP-1 macrophages. Following 18 h serum starvation, cells were re-fed with serum-replete media and either treated immediately with/without MSU crystals (500 μg/mL) and heme (30 μM)), SR8278 (1 μM), heme+SR8278 or vehicle (0.1M NaOH + 0.1% DMSO) for 12 h (0–12 h timepoint) or rested for 12 h and then media-changed to fresh serum-replete media with/without MSU crystals and heme/SR8278/vehicle (12–24 h timepoint). (A) Caspase-1 activity normalized to cell number as measured at the 0–12 h timepoint and (B) 12–24 h timepoint. (C) Levels of secreted IL-1β measured by ELISA at the 0–12 h and (D) 12–24 h timepoint. To ensure all cells were exposed to the same vehicles, SR8278 vehicle (0.1%) DMSO) was added to the heme-only treatment and heme vehicle (0.1M NaOH) added to the SR8278-only treatment. Data shown are mean ± SEM for 3 experimental replicates. All data were analyzed by one-way ANOVA (post-hoc Tukey) p < .05 was considered statistically significant.

Article Snippet: Secreted IL- 1β protein levels were measured in the cell culture supernatant using a Duoset Human IL- 1β ELISA kit (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Representative fluorescence microscopy images of mesenchymal stromal cells (MSCs) infected by IL-4 and/or PDGF-BB lentiviral vectors. GFP-MSCs, MSCs infected with control GFP-positive lentivirus vector; IL4-MSCs, MSCs infected with rIL-4 secreting GFP-positive lentivirus vector; PDGF-BB-MSCs, MSCs infected with hPDGF-BB secreting GFP-positive lentivirus vector; IL4-PDGF-BB-MSCs, previously established IL4-MSCs infected with hPDGF-BB secreting RFP-positive lentivirus vector.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Fluorescence, Microscopy, Infection, Control, Plasmid Preparation

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: The experimental outline of in vitro experiments. GFP-MSCs, IL4-MSCs, PDGF-BB-MSCs, or IL4-PDGF-BB-MSCs were seeded into T75 flasks and cultured in an MSC growth medium for 1 day. For the non-preconditioning MSC groups, the cells were cultured for 3 days with a fresh MSC growth medium. For the preconditioning MSC groups, cells were cultured in the MSC preconditioning medium for 3 days. The cells from all eight groups were washed three times with Dulbecco’s phosphate-buffered saline. The cells were trypsinized and seeded for each experiment, including cell proliferation, IL4 and PDGF-BB expression level, and osteogenic differentiation performed by alkaline phosphatase (ALP) staining and Alizarin red-staining. Abbreviations: MSCs, mesenchymal stromal cells; pMSCs, preconditioning MSCs; TNFα, tumor necrosis factor-alpha; LPS, lipopolysaccharide.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: In Vitro, Cell Culture, Saline, Expressing, Staining

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Fold change in the amount of dsDNA compared to day 1.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques:

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: ( A ) IL4 and PDGF-BB expression levels on day 3 in the IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups. IL4 expression levels on day 3 in the IL4-MSC, IL4-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 77.7 ± 4.4 ng/mL, 34.6 ± 5.7 ng/mL, 16.8 ± 9.4 ng/mL, and 41.8 ± 5.0 ng/mL, respectively. PDGF-BB expression levels on day 3 in the PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 121.7 ± 20.1 pg/mL, 50.3 ± 20.4 pg/mL, 8.7 ± 2.0 pg/mL, 71.5 ± 7.8 pg/mL, respectively. ( B ) IL4 and PDGF-BB expression level-dsDNA ratios on day 3 in the IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups. IL4 expression level-dsDNA ratios on day 3 in the IL4-MSC, IL4-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 7.3 ± 0.6, 1.8 ± 0.2, 1.8 ± 1.0, and 2.3 ± 0.1, respectively. PDGF-BB expression level-dsDNA ratios on day 3 in the PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 5.6 ± 0.9, 3.7 ± 1.4, 1.0 ± 0.3, and 3.9 ± 0.1, respectively.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Expressing

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Alkaline phosphatase-staining in all eight groups 2 weeks after seeding. The percentages of stained area in the GFP-MSC, GFP-pMSC, IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 19.8 ± 5.3%, 28.3 ± 1.4%, 4.3 ± 0.7%, 6.0 ± 2.4%, 44.2 ± 11.7%, 42.5 ± 8.1%, 21.4 ± 1.5%, and 25.7 ± 1.0%, respectively. Abbreviations: ALP, alkaline phosphatase.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Staining

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Alizarin red-staining in all eight groups 4 weeks after seeding. The percentages of stained area in the GFP-MSC, GFP-pMSC, IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 33.1 ± 4.7%, 45.1 ± 4.9%, 0.64 ± 0.08%, 0.58 ± 0.03%, 51.7 ± 14.5%, 40.7 ± 6.7%, 17.3 ± 2.5%, and 41.2 ± 1.7%, respectively.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Staining

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: The summary of the results.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Expressing

Journal: Nutrients

Article Title: Effect of Fermented Soybean (FSB) Supplementation on Gastroesophageal Reflux Disease (GERD)

doi: 10.3390/nu16162779

Figure Lengend Snippet: Changes in inflammation levels of participant during study, values are expressed as mean ± SD.

Article Snippet: Interleukin-4 (IL-4) levels were assayed using Elabscience Human IL-4 ELISA Kit (Elabscience Biotechnology Co., Ltd., Houston, TX, USA), Interleukin-6 (IL-6) levels were assayed using Elabscience Human IL-6 ELISA Kit (Elabscience Biotechnology Co., Ltd., Houston, TX, USA), and Interleukin-8 (IL-8) levels were assayed using Elabscience Human IL-8 ELISA Kit (Elabscience Biotechnology Co., Ltd., Houston, TX, USA).

Techniques: