Journal: European Journal of Medical Research

Article Title: Detection of skin α-synuclein using RT-QuIC as a diagnostic biomarker for Parkinson’s disease in the Chinese population

doi: 10.1186/s40001-024-01705-x

Figure Lengend Snippet: Detection of α-Syn seeding activity by skin RT-QuIC assay in patients with PD and controls. A RT-QuIC detection of α-Syn seeding activity in the cervical skin homogenates from 4 patients with PD and 4 controls. B RT-QuIC detection of α-Syn seeding activity in the skin homogenates of 30 patients and 28 controls. C Scatter graph of maximal thioflavin T (ThT) fluorescence at 70 h of the α-Syn seeding activity by RT-QuIC assay. D ROC curve and AUC calculation for comparison of α-Syn seeding activity between patients with PD and controls ( P < 0.001)

Article Snippet: Each reaction consisted of 2 μL of skin homogenate (0.245–0.5 μg/μL) as a seed and 98 μL of α-Syn RT-QuIC reaction mixture with six 0.8 mm silica beads (OPS Diagnostics, Lebanon, NJ) per well in a 96-well plate (Nalgene Nunc International, 265301).

Techniques: Activity Assay, Fluorescence, Comparison

Journal: European Journal of Medical Research

Article Title: Detection of skin α-synuclein using RT-QuIC as a diagnostic biomarker for Parkinson’s disease in the Chinese population

doi: 10.1186/s40001-024-01705-x

Figure Lengend Snippet: Detection of fibrillary α-Syn in the RT-QuIC product of PD patients. A , B The structural features of α-Syn aggregates extracted from PD-seeded RT-QuIC reactions were analyzed by employing transmission electron microscopy (TEM) and atomic force microscopy (AFM), which could not be observed in the control samples. C Congo red staining, observed under polarized light, exhibited an apple-green color in PD patients’ samples obtained from the RT-QuIC assay, whereas the control samples without PD showed a negative result. The scale bar represents 200 μm. D Analysis of Dot Blot both T0 and RT-QuIC Reaction in PD and Control Groups: Dot blot analysis was conducted on skin samples from three PD and three controls at both T0 and after RT-QuIC reactions using α-Syn filament conformation-specific antibodies (top panel) and total α-Syn antibodies (bottom panel). Densitometric quantification of α-Syn filament conformation-specific and total α-Syn levels revealed significantly increased levels of α-Syn filaments for the PD group compared to the control group, while total αSyn levels did not differ between groups. Results were analyzed using Student’s two-sample t -test. ** p < 0.01 and not significant (ns) when P > 0.05

Article Snippet: Each reaction consisted of 2 μL of skin homogenate (0.245–0.5 μg/μL) as a seed and 98 μL of α-Syn RT-QuIC reaction mixture with six 0.8 mm silica beads (OPS Diagnostics, Lebanon, NJ) per well in a 96-well plate (Nalgene Nunc International, 265301).

Techniques: Transmission Assay, Electron Microscopy, Microscopy, Staining, Dot Blot

Journal: European Journal of Medical Research

Article Title: Detection of skin α-synuclein using RT-QuIC as a diagnostic biomarker for Parkinson’s disease in the Chinese population

doi: 10.1186/s40001-024-01705-x

Figure Lengend Snippet: Skin RT-QuIC results exhibit no correlation with disease progression. Raw RT-QuIC assay parameters reveal no association with PD duration: longer disease duration ( A , E ), higher Hoehn and Yahr (H&Y) stage ( B , F ), higher Unified Parkinson’s Disease Rating Scale (UPDRS) score ( C , G ), and lower Mini-Mental State Examination (MMSE) score ( D , H ) do not exhibit a higher final ThT fluorescence or longer lag phase

Article Snippet: Each reaction consisted of 2 μL of skin homogenate (0.245–0.5 μg/μL) as a seed and 98 μL of α-Syn RT-QuIC reaction mixture with six 0.8 mm silica beads (OPS Diagnostics, Lebanon, NJ) per well in a 96-well plate (Nalgene Nunc International, 265301).

Techniques: Fluorescence