Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular sirolimus concentration is reduced by tacrolimus in human pancreatic islets in vitro.

doi: 10.1111/tri.12617

Figure Lengend Snippet: Figure 1 Intracellular concentrations of TAC in human islets. Human islets were cultured with TAC (10 or 30 lg/l), SRL (10 or 30 lg/l), or the combi- nation thereof for 24–48 h before the intracellular concentration of TAC was measured in islet lysate and normalized to total protein as detailed in methods. Data are presented as the mean SD, n = 6 for each group. TAC, tacrolimus; SRL, sirolimus; *P < 0.04; ** P < 0.007; *** P < 0.0006.

Article Snippet: The islets were exposed to 10 and 30 lg/l of tacrolimus (Santa Cruz Biotechnology, Dallas, TX, USA) or sirolimus (Toronto Research Chemicals, Toronto, Ontario, Canada) or the combination thereof for 24 h and 48 h at 37 °C (5% CO2).

Techniques: Cell Culture, Concentration Assay

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular sirolimus concentration is reduced by tacrolimus in human pancreatic islets in vitro.

doi: 10.1111/tri.12617

Figure Lengend Snippet: Figure 2 Intracellular concentrations of SRL in human islets. Human islets were cultured with TAC (10 or 30 lg/l), SRL (10 or 30 lg/l), or the combi- nation thereof for 24–48 h before the intracellular concentration of SRL was measured in islet lysate and normalized to total protein as detailed in methods. Data are presented as the mean SD, n = 6 for each group. TAC: tacrolimus; SRL: sirolimus; *** P < 0.001; **** P < 0.0001.

Article Snippet: The islets were exposed to 10 and 30 lg/l of tacrolimus (Santa Cruz Biotechnology, Dallas, TX, USA) or sirolimus (Toronto Research Chemicals, Toronto, Ontario, Canada) or the combination thereof for 24 h and 48 h at 37 °C (5% CO2).

Techniques: Cell Culture, Concentration Assay

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular sirolimus concentration is reduced by tacrolimus in human pancreatic islets in vitro.

doi: 10.1111/tri.12617

Figure Lengend Snippet: Figure 3 Effect of CsA on intracellular concentration of SRL in human islets. Human islets were cultured with the combination of SRL (30 lg/l) and CsA (5 lg/ml), or the drug alone for 24 h before the intracellular concentration of SRL (a) or CsA (b) was measured in islet lysate and nor- malized to total protein as detailed in methods. Data are calculated as percentages of control and are presented as mean SD, n = 6 for each group. CsA, cyclosporine A; SRL, sirolimus.

Article Snippet: The islets were exposed to 10 and 30 lg/l of tacrolimus (Santa Cruz Biotechnology, Dallas, TX, USA) or sirolimus (Toronto Research Chemicals, Toronto, Ontario, Canada) or the combination thereof for 24 h and 48 h at 37 °C (5% CO2).

Techniques: Concentration Assay, Cell Culture, Control

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular sirolimus concentration is reduced by tacrolimus in human pancreatic islets in vitro.

doi: 10.1111/tri.12617

Figure Lengend Snippet: Figure 4 The effect of SRL, TAC, or CsA on phosphorylation of p70S6k in islets. Human islets were cultured with TAC (30 lg/l), SRL (30 lg/l), or the combination thereof for 24 h before the presence of p- p70s6k was assessed by the cell-signaling Bio-Plex assay in human islet cell lysate and normalized to total protein (a). In a parallel experiment, human islets were cultured with SRL (30 lg/l), CsA (5 lg/ml), or the combination thereof for 24 h before p-p70S6k was detected in the lysate and normalized to total protein. Data are calculated as ratio to control and are presented as mean SD, n = 3–6 for each group. TAC, tacrolimus; SRL, sirolimus; CsA, cyclosporine A, ** P < 0.01, **** P < 0.0001.

Article Snippet: The islets were exposed to 10 and 30 lg/l of tacrolimus (Santa Cruz Biotechnology, Dallas, TX, USA) or sirolimus (Toronto Research Chemicals, Toronto, Ontario, Canada) or the combination thereof for 24 h and 48 h at 37 °C (5% CO2).

Techniques: Phospho-proteomics, Cell Culture, Plex Assay, Control

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular sirolimus concentration is reduced by tacrolimus in human pancreatic islets in vitro.

doi: 10.1111/tri.12617

Figure Lengend Snippet: Figure 5 Oxygen consumption rates (OCR) in human islets after treatment of TAC, SIR, or SIR+TAC. Human islets were cultured with TAC (30 lg/l), SIR (30 lg/l), or a combination thereof for 24 h before the glucose-stimulated OCR was measured as indicated in methods. OCR is expressed as percentage of baseline and is presented as the mean SD, n = 6 for each group. TAC, tacrolimus; SRL, sirolimus, ** P < 0.01.

Article Snippet: The islets were exposed to 10 and 30 lg/l of tacrolimus (Santa Cruz Biotechnology, Dallas, TX, USA) or sirolimus (Toronto Research Chemicals, Toronto, Ontario, Canada) or the combination thereof for 24 h and 48 h at 37 °C (5% CO2).

Techniques: Cell Culture

Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular sirolimus concentration is reduced by tacrolimus in human pancreatic islets in vitro.

doi: 10.1111/tri.12617

Figure Lengend Snippet: Figure 6 Expression of ABCB1 (Pgp), OATP1B1, and CYP3A4 in human islets. Human islets were cultured for 24 h before the expression of the drug transporter (ABCB1(Pgp) and OATP1B1), and the metabolic enzyme CYP3A4 was evaluated. RNA was prepared and subjected to qPCR as detailed in methods. The reference gene index is calculated by the mean of ALAS1, B2M, and RPL13A expression and used to normalize the expression of target genes in isolated hepatocytes relative to the mRNA level of ABCB1(Pgp), OATP1B1, and CYP3A4 in human islets (a). OATB1 mRNA expression in human islets was normalized to reference gene index after exposure to TAC (30 lg/l), SRL (30 lg/l), or a combination thereof for 24 h (b). Represen- tative immunofluorescence image of dispersed human islets stained for insulin (green), ABCB1(Pgp) (red) and nuclear staining with DAPI (blue) (c), or glucagon (green), ABCB1(Pgp) (red) and nuclear staining wit DAPI (blue) (d). Data are presented as mean SD, n = 4–5 for each group. TAC, tacroli- mus; SRL, sirolimus; *P < 0.05; ** P < 0.01.

Article Snippet: The islets were exposed to 10 and 30 lg/l of tacrolimus (Santa Cruz Biotechnology, Dallas, TX, USA) or sirolimus (Toronto Research Chemicals, Toronto, Ontario, Canada) or the combination thereof for 24 h and 48 h at 37 °C (5% CO2).

Techniques: Expressing, Cell Culture, Isolation, Staining

Journal: Cancer Management and Research

Article Title: MIR31HG Expression Predicts Poor Prognosis and Promotes Colorectal Cancer Progression

doi: 10.2147/CMAR.S351928

Figure Lengend Snippet: Association between MIR31HG expression and related genes. ( A ) Relationship between MIR31HG and the related genes were shown in a heatmap. ( B ) The expression correlation between MIR31HG and the selected genes were analyzed. ( C ) Representative images of immunohistochemistry showing MIR31HG-associated genes for distinct pathologic stages in colorectal cancer tissues (n=5 for each group). Scale bars, 50 μm. *P<0.05, **P<0.01 compared with pathologic stage I in colorectal cancer tissues. ( D ) Western blot analysis was performed to determine the expression of TPBG, FYN, ETV1 and TRERF1. ( E ) The expression of select genes was quantified by qRT-PCR in MIR31HG-silenced HT-29 cells. *P<0.05, **P<0.01 compared with negative control.

Article Snippet: The following antibodies were used: anti-TPBG antibody (Proteintech), anti-FYN antibody (Proteintech), anti-ETV1 antibody (Abcam) and anti-TRERF1 antibody (Proteintech).

Techniques: Expressing, Immunohistochemistry, Western Blot, Quantitative RT-PCR, Negative Control

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa and CsA on the expression of DCs surface markers. Immature DCs were differentiated in the environment of immunosuppressive agents: Rapa (Rapa-DC) or CsA (CsA-DC) and without drugs (DC). Gating strategy for flow cytometric analysis of DCs: a time gate was initially applied to exclude any electronic noise and artifact (not shown here). Next, based on size and granularity, DCs were gated in a forward scatter area (FSC-A) versus side scatter area (SSC-A) plot. Then, doublet cells were excluded using FSC-A/FSC-height (FSC-H) parameters. Within the singlet cell population CD11c + DCs were gated ( A ), followed by expression of individual markers shown on representative histograms for DC, Rapa-DC and CsA-DC ( B ). The averages of the percentage of positively labelled CD11c + DCs and mean fluorescence intensity in bracket 7–14 different donors are reported on each histogram. The white shade indicates FMO control. Representative dot plots are presented

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Fluorescence, Control

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa and CsA on the cytokine production by immature DCs. Immature DCs were differentiated in the environment of immunosuppressive agents: Rapa (Rapa-DC) or CsA (CsA-DC) and without drugs (DC). Cytokine expression was measured by intracellular staining (IL-4; A ) or by ELISA of culture supernatants (IL-10, TGF-β and IL-6; B – D , respectively). Results are the averages ± SD of the percentage of positively labelled CD11c + DCs ( A ; n = 5) or cytokine secretion ( B – D ; n = 19). ** p ≤ 0.01; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa and CsA on phagocytic capacity of DCs. Immature DCs were differentiated in the environment of immunosuppressive agents: Rapa (Rapa-DC) or CsA (CsA-DC) and without drugs (DC). Cellular FITC-dextran uptake ( A , B ; n = 7) and the expression of phagocytosis receptors: CD36 ( C , D ; n = 10) and DEC205 ( F , G ; n = 11) were determined by flow cytometry. Results are the averages ± SD of the percentage of positively labelled CD11c + DCs and mean fluorescence intensity (MFI). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa and CsA on the expression of surface markers on LPS-activated DCs

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Marker, Control, Activation Assay

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa and CsA on the cytokine production by LPS-activated DCs. Immature DCs generated in the environment of: Rapa (Rapa-DC + LPS), CsA (CsA-DC + LPS) and without drugs (DC + LPS) were activated with LPS. Additionally, DCs, which differentiated without drugs, were activated with LPS simultaneously with Rapa (DC + LPS + Rapa) or CsA (DC + LPS + CsA). Cytokine expression was measured by intracellular staining (IL-4; A ) or by ELISA of culture supernatants (IL-10, TGF-β and IL-6; B – D , respectively). Results are the averages ± SD of the percentage of positively labelled CD11c + DCs ( A ) or cytokine secretion ( B – D ) from at least five different donors. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Generated, Expressing, Staining, Enzyme-linked Immunosorbent Assay

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa and CsA on PD-L1 expression on DCs. Immature DCs were differentiated in the environment of immunosuppressive agents: Rapa (Rapa-DC) or CsA (CsA-DC) and without drugs (DC). Expression of PD-L1 was determined by flow cytometry. Results are the averages ± SD of the percentage of CD11c + PD-L1 + DCs ( A ) and PD-L1 mean fluorescence intensity (MFI; B ) from nine different donors. * p ≤ 0.05; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa-DC and CsA-DC on the expression of CD95 on CD4 + and CD8 + T cells. Immature DCs, Rapa-DC and CsA-DC were cocultured with T cells. Expression of CD95 was determined by flow cytometry. Results are the averages ± SD of the percentage of positively labelled CD4 + ( A ) or CD8 + ( C ) T cells and mean fluorescence intensity (MFI) for CD4 + ( B ) or CD8 + ( D ) T cells, from ten different donors. * p ≤ 0.05, ** p ≤ 0.01; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa-DC and CsA-DC on T regulatory cells populations. Immature DCs, Rapa-DC and CsA-DC were cocultured with T cells. The percentage of CD4 + CD25 high Foxp3 + ( A ), CD8 + CD25 + CD28 + ( B ) and CD8 + CD25 – CD28 – ( C ) T cells were determined by flow cytometry. Results are the averages ± SD from ten different donors. * p ≤ 0.05; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Flow Cytometry

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms

doi: 10.1007/s00005-021-00632-7

Figure Lengend Snippet: Effects of Rapa-DC and CsA-DC on the production of Th1- and Th2-characteristic cytokines. Immature DCs, Rapa-DC and CsA-DC were cocultured with T cells. Cytokine expression was measured by ELISA of culture supernatants for the presence of Th1 cytokines: IL-2 ( A ), IFN-γ ( B ) and Th2 cytokines: IL-4 ( C ), IL-5 ( D ), IL-10 ( E ), IL-13 ( F ). Results are the averages ± SD of cytokine secretion from 19 different donors. * p ≤ 0.05, ** p ≤ 0.01; p values were calculated by Wilcoxon matched pair test

Article Snippet: To generate immature DCs with tolerogenic properties, 200 ng/ml CsA (CsA-DC; Novartis Pharma, Germany) or 20 ng/ml Rapa (Rapa-DC; Wyeth-Lederle Pharma, UK) was added from the beginning of cultures.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: Bioactive Materials Facilitate the Restoration of Neurological Function Post Cerebral Ischemic Stroke

doi: 10.2147/IJN.S493987

Figure Lengend Snippet: Polysaccharide sulfate-based nanocarriers deliver targeted neuroprotective agent rapamycin in the management of cerebral infarction. ( A ) The schematic design of RAPA @ tRPCS. ( B ) Phenotypic changes in microglia upon exposure to various nanoparticles (n = 3; * P < 0.05, *** P < 0.005). ( C ) The effect of different nanoparticles on microglia size (n = 3; * P < 0.05, ** P < 0.01, **** P < 0.001). ( D ) The infarct volumes at 7 days after tMCAO were measured with ImageJ in different groups (n = 3; ** P < 0.01, *** P < 0.005, **** P < 0.001). Reprinted with permission from Cao Y, Yu Y, Pan L, et al. Sulfated polysaccharide-based nanocarrier drives microenvironment-mediated cerebral neurovascular remodeling for ischemic stroke treatment. Nano Lett. 2024;24(17):5214–5223. Copyright 2024, American Chemical Society.

Article Snippet: , , Targeted delivery of RAPA , Sulfate polysaccharide nanocarrier loaded with RAPA , Murine model of transient MCAO , [ ] .

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Bioactive Materials Facilitate the Restoration of Neurological Function Post Cerebral Ischemic Stroke

doi: 10.2147/IJN.S493987

Figure Lengend Snippet: Conspectus of Anti-Ischemia Tactics and Repair Mechanisms in Management of Ischemic Stroke

Article Snippet: , , Targeted delivery of RAPA , Sulfate polysaccharide nanocarrier loaded with RAPA , Murine model of transient MCAO , [ ] .

Techniques: Membrane, Activity Assay, Recombinant, Mouse Assay, Activation Assay, Translocation Assay, Expressing, Migration