Journal: Journal of Nanobiotechnology

Article Title: Peripheral administration of nanomicelle-encapsulated anti-Aβ oligomer fragment antibody reduces various toxic Aβ species in the brain

doi: 10.1186/s12951-023-01772-y

Figure Lengend Snippet: Quantitatively evaluation of immune-positivity of toxic Aβs in brain sections. After administering phosphate-buffered saline (PBS) in A anti-amyloid β oligomer (AβO) 6H4 antibody fragments (Fabs) (6H4 Fab PM) and B anti-Aβ 3D6 Fabs encapsulated in PMs (3D6 Fab PM), C anti-AβO 6H4 Fabs (6H4 Fab), and D Alzheimer’s disease (AD) model and E untreated wild-type (WT) mice, immunohistochemistry with anti-Aβ 82E1 antibody was performed. Panels A–E reveal significantly narrower plaque diameters F and smaller 82E1positive areas G in the 6H4 Fab PM group (black bars) than in the PBS group (white bars) . Images of immunofluorescent positive signals for anti-AβO 6H4 antibody (magenta) and thioflavin S (green) in AD mice treated with PBS ( H ), 6H4 Fab PMs ( I ), 3D6 Fab PM ( J ), and 6H4 Fab ( K ), and WT mice (L). Quantitative analysis of the % region of interest (ROI) for 6H4 antibody ( M ) and thioflavin S ( N ) positive signals of AD mice observed in panels H–L revealed significantly reduced %ROI and thioflavin S-positive signals in the 6H4 Fab PM group compared with that in the PBS group . Immunoreactive-images for anti-N3pE Aβ antibody in AD mice treated with PBS ( O ) , 6H4 Fab PM ( P ), 3D6 Fab PM ( Q ) , and 6H4 Fab ( R ) and WT mice ( S ). Enlarged views are presented inside dashed lines on each figure. N3pE Aβ-positive Aβ plaques in the 6H4 Fab PM group showed small diffuse-like plaques without Aβ cores ( P ). Quantitative values of the immunostained N3pE Aβs represent the average numbers of plaques in 10 figures in panels O–S. The 6H4 Fab PM group showed significantly smaller N3pE Aβ antibody-positive areas ( T ) and diameters ( U ) than those in the PBS group. Scale bar = 100 μm. One-way analysis of variance with Tukey’s post-hoc test was performed. Values = mean ± standard error of the mean. * p < 0.05, ** p < 0.01

Article Snippet: Next, the sections were incubated with filtered 0.002% thioflavin S (Merck KGaA, Darmstadt, Germany) solution dissolved in PBS for 10 min.

Techniques: Immunohistochemistry

Journal: Journal of Nanobiotechnology

Article Title: Peripheral administration of nanomicelle-encapsulated anti-Aβ oligomer fragment antibody reduces various toxic Aβ species in the brain

doi: 10.1186/s12951-023-01772-y

Figure Lengend Snippet: ThT assay of Aβ aggregation. The inhibitory effects of anti-amyloid β (Aβ)O antibody 6H4, anti-Aβ antibody 3D6, and anti-Aβ aggregate-specific antibody GTX134510 on Aβ42 aggregation were evaluated using a thioflavin T (ThT) assay. A Aβ alone (white circles) continued to increase the self-aggregation rate even after 1 h of incubation with ThT. In contrast, 6H4 (black circles) showed stable inhibition of Aβ42 aggregation throughout the incubation period. However, compared with 6H4, GTX 134,510 (black triangles) showed a significantly decreased inhibition effect for approximately 3 h of incubation; 3D6 (white squares) showed a significantly decreased inhibition effect after approximately 8 h of incubation. B Compared with Aβ alone (white bar) and GTX134510 (hatched bar), 6H4 (black bar) showed a significant inhibition of the Aβ42 aggregation rate after 6 h of incubation. C Compared with Aβ alone, 6H4 maintained a 99% inhibition rate on Aβ42 aggregation even after 12 h of incubation; this was significantly higher than the inhibition rate maintained by 3D6 (horizontal striped bar). Values are expressed as mean ± standard error of the mean. Statistical evaluations were performed using one-way or two-way analysis of variance with a Tukey’s post-hoc test. ** p < 0.01

Article Snippet: Next, the sections were incubated with filtered 0.002% thioflavin S (Merck KGaA, Darmstadt, Germany) solution dissolved in PBS for 10 min.

Techniques: ThT Assay, Incubation, Inhibition

Journal: ACS Omega

Article Title: Substituent, Charge, and Size Effects on the Fluorogenic Performance of Amyloid Ligands: A Small-Library Screening Study

doi: 10.1021/acsomega.7b00231

Figure Lengend Snippet: Binding Constants ( K d ) of OPE1 to Lysozyme and Insulin Amyloid. K d Values of Thioflavin T are Included as a Reference

Article Snippet: Thioflavin T (ThT) was obtained from VWR International (Radnor, PA).

Techniques: Binding Assay