Journal: Journal of extracellular biology

Article Title: Single vesicle analysis of aqueous humor in pediatric ocular diseases reveals eye specific CD63-dominant subpopulations

doi: 10.1002/jex2.36

Figure Lengend Snippet: Tetraspanin expression analysis of unprocessed aqueous humor samples using the SP-IRIS analysis. Samples were analysed by the Exoview R100 system. (a) Representative fluorescent images detected by fluorescent-conjugated antibodies (red: CD63-AF647, green: CD81-AF555 and blue: CD9-AF488) on Exoview tetraspanin chip immunocapture spots. (a) Total fluorescent particle counts normalized to isotype control IgG on a single capture spot in individual samples. (c,d) Total fluorescent particle counts normalized to isotype control IgG determined by detection antibody on three capture spots in five pediatric disease states. Error bars represent ± one S.E.M

Article Snippet: The chips were then washed three times in solution A (ExoView Human Tetraspanin Kit, NanoView Biosciences, USA) and followed by incubation with Immunocapture antibodies that consist of anti-CD9 CF488, anti-CD81 CF555, and anti-CD63 CF647.

Techniques: Expressing, Control

Journal: Journal of extracellular biology

Article Title: Single vesicle analysis of aqueous humor in pediatric ocular diseases reveals eye specific CD63-dominant subpopulations

doi: 10.1002/jex2.36

Figure Lengend Snippet: Colocalization analysis of tetraspanin subgroups of unprocessed aqueous humor samples using the SP-IRIS analysis. Samples were analysed by the Exoview R100 system. (a) Representative fluorescent images detected by fluorescent-conjugated antibodies (red: CD63-AF647, green: CD81-AF555 and blue: CD9-AF488) on one immunocapture spot. (b,c) Tetraspanin subpopulation distribution in five pediatric disease states. Data were denoted as normalized fluorescent particle counts. (d–f) Percentages of tetraspanin subpopulation distribution in five pediatric disease states. Error bars represent ± one S.E.M. * p < 0.05, ** p < 0.01 calculated by Mann-Whitney U test

Article Snippet: The chips were then washed three times in solution A (ExoView Human Tetraspanin Kit, NanoView Biosciences, USA) and followed by incubation with Immunocapture antibodies that consist of anti-CD9 CF488, anti-CD81 CF555, and anti-CD63 CF647.

Techniques: MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Comparison between Polybutylcyanoacrylate Nanoparticles with Either Surface-Adsorbed or Encapsulated Brain-Derived Neurotrophic Factor on the Neural Differentiation of iPSCs

doi: 10.3390/ijms20010182

Figure Lengend Snippet: The average diameter (Dav), zeta potential, polydispersity index (PDI), and loading efficiencies of BDNF and SPIO for the four types of nanoparticles.

Article Snippet: The free BDNF in solution was analyzed with a human BDNF enzyme linked immunosorbent assay (ELISA) kit (R&D Systems) after ultrafiltration.

Techniques: Zeta Potential Analyzer

Journal: International Journal of Molecular Sciences

Article Title: Comparison between Polybutylcyanoacrylate Nanoparticles with Either Surface-Adsorbed or Encapsulated Brain-Derived Neurotrophic Factor on the Neural Differentiation of iPSCs

doi: 10.3390/ijms20010182

Figure Lengend Snippet: Transmission electron microscope (TEM) images of the nanoparticles at ×200,000 magnification, showing that the nanoparticles have a dense core and an ill-defined halo that is visible for those that contain the surface-adsorbed T80 or BDNF.

Article Snippet: The free BDNF in solution was analyzed with a human BDNF enzyme linked immunosorbent assay (ELISA) kit (R&D Systems) after ultrafiltration.

Techniques: Transmission Assay, Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Comparison between Polybutylcyanoacrylate Nanoparticles with Either Surface-Adsorbed or Encapsulated Brain-Derived Neurotrophic Factor on the Neural Differentiation of iPSCs

doi: 10.3390/ijms20010182

Figure Lengend Snippet: Cell viability determined by the XTT assay for the different PBCA NPs at the dose range of 5 to 100 μg/mL on ( a ) rat brain microvascular endothelial cells (RBMECs), ( b ) rat astrocyte (CTX TNA2), and ( c ) iPSCs. A concentration-dependent cytotoxicity profile is evident, and a concentration of up to 25 μg/mL appears safe for the RBMECs and CTX TNA2, and up to 50 μg/mL for the iPSCs. No significant differences are found in between the adsorption and encapsulation system of BDNF with and without T80 coating across the cell lines, except for the iPSCs in which T80 appears cytotoxic at a high nanoparticle concentration of 100 μg/mL. * p < 0.05 to control, # p < 0.001.

Article Snippet: The free BDNF in solution was analyzed with a human BDNF enzyme linked immunosorbent assay (ELISA) kit (R&D Systems) after ultrafiltration.

Techniques: XTT Assay, Concentration Assay, Adsorption, Encapsulation, Control

Journal: International Journal of Molecular Sciences

Article Title: Comparison between Polybutylcyanoacrylate Nanoparticles with Either Surface-Adsorbed or Encapsulated Brain-Derived Neurotrophic Factor on the Neural Differentiation of iPSCs

doi: 10.3390/ijms20010182

Figure Lengend Snippet: ( a ) Immunofluorescence staining at Day 7 for BDNF (green) of the iPSCs exposed to 125 pg/mL of BDNF alone or 25 μg/mL of nanoparticles with or without BDNF, and ( b ) quantification of BDNF expression by the normalized fluorescence intensity; the levels of BDNF expression appear similar for the control, PBCA-SPIO NPs, and free BDNF, where significant increases can be found for the BDNF-containing nanoparticles, which is associated with a greater dispersion of the cell cluster and elongated cell shape. * p < 0.05 to control.

Article Snippet: The free BDNF in solution was analyzed with a human BDNF enzyme linked immunosorbent assay (ELISA) kit (R&D Systems) after ultrafiltration.

Techniques: Immunofluorescence, Staining, Expressing, Fluorescence, Control, Dispersion

Journal: International Journal of Molecular Sciences

Article Title: Comparison between Polybutylcyanoacrylate Nanoparticles with Either Surface-Adsorbed or Encapsulated Brain-Derived Neurotrophic Factor on the Neural Differentiation of iPSCs

doi: 10.3390/ijms20010182

Figure Lengend Snippet: Immunofluorescence staining at Day 7 for ( a ) nestin (red) and the neurofilament-heavy chain (NF H) (green), and ( b ) beta III tubulin (green) on the iPSCs exposed to 125 pg/mL of free BDNF or 25 μg/mL of nanoparticles with or without BDNF. The control, PBCA-SPIO NPs and free BDNF, show a limited number of nestin and NF H-positive cells; by contrast, positively-stained cells are easily identifiable when the BDNF delivery systems are used. Similarly, the control and PBCA-SPIO NP show very few numbers of beta III tubulin-positive cells, and although it is higher with free BDNF, neural morphology cannot be found. Neural differentiation is clearly seen in cells treated with the BDNF-containing nanoparticles; furthermore, surface-adsorbed BDNF is associated with a greater degree of axon elongation and intercellular connections. ( c ) Flow cytometry analysis of beta III tubulin-positive cells on iPSCs exposed to free BDNF or nanoparticles; the control and PBCA-SPIO NPs show similar percentages. This percentage increases to 31.4 ± 2.7% with free BDNF, and further increases to 41.9 ± 3.4% with the BDNF delivery systems.

Article Snippet: The free BDNF in solution was analyzed with a human BDNF enzyme linked immunosorbent assay (ELISA) kit (R&D Systems) after ultrafiltration.

Techniques: Immunofluorescence, Staining, Control, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Comparison between Polybutylcyanoacrylate Nanoparticles with Either Surface-Adsorbed or Encapsulated Brain-Derived Neurotrophic Factor on the Neural Differentiation of iPSCs

doi: 10.3390/ijms20010182

Figure Lengend Snippet: ( a ) The transendothelial electrical resistance (TEER) (Ωcm 2 ) across the contact co-culture system of RBMECs and CTX TNA2 for the BDNF-containing PBCA-SPIO NPs at 25 μg/mL for 4 h. The TEER of the treated cells are significantly lower than the control, and the values of the adsorption and the encapsulation system of BDNF only differ with respect to the presence and absence of T80 coating, which, on average, measures 382.9 ± 18.1 and 426.9 ± 19.7 Ωcm 2 , respectively. ( b ) The permeabilities of the nanoparticles (cm/s) correspond to the degree of reduction in the TEERs, which are significantly higher with T80 coating regardless of the physical location of BDNF. * p < 0.05 to control, # p < 0.05.

Article Snippet: The free BDNF in solution was analyzed with a human BDNF enzyme linked immunosorbent assay (ELISA) kit (R&D Systems) after ultrafiltration.

Techniques: Co-Culture Assay, Control, Adsorption, Encapsulation