Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Measurement of total and pSer129-aSyn levels in detergent-insoluble brain fractions by AlphaLISA. ( a and b ) total and pSer129-aSyn levels in PD and control (C) groups. ( c and d ) total and pSer129-aSyn levels across controls, EOPD and LOPD. ( e and f ) Comparison of total and pSer129-aSyn levels in FP-LO and SP-LO. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons (c and d) and the Mann–Whitney U test for pairwise comparisons (a, b, c and e), and Student’s t-test (f)

Article Snippet: The reaction mixture contained 1X phosphate-buffered saline (KD Medical, PBS 10X, pH 8.0), 0.0006% sodium dodecyl sulfate (SDS), 10 μM Thioflavin T (ThT), and 0.1 mg/mL recombinant human aSyn monomer (Proteos, RP-010).

Techniques: Control, Comparison, MANN-WHITNEY

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: aSyn seeding bioactivity measured by FRET FC assay. ( a ) Comparison of aSyn seeding activity in PD and control (C) groups. ( b ) Comparison of aSyn seeding activity in EOPD and LOPD disease subgroups. ( c ) Comparison of aSyn seeding activity in FP-LO and SP-LO subgroups. ( d ) Correlation between pSer129-aSyn levels and FRET seeding bioactivity in PD cases. ( e ) Cell viability analysis following treatment with detergent-insoluble brain fractions. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons and the Mann–Whitney U test for pairwise comparisons. For the correlation analysis, a two-tailed Spearman’s rank correlation test was used, and r and p values are indicated on the plot

Article Snippet: The reaction mixture contained 1X phosphate-buffered saline (KD Medical, PBS 10X, pH 8.0), 0.0006% sodium dodecyl sulfate (SDS), 10 μM Thioflavin T (ThT), and 0.1 mg/mL recombinant human aSyn monomer (Proteos, RP-010).

Techniques: Comparison, Activity Assay, Control, MANN-WHITNEY, Two Tailed Test

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Intracellular aSyn aggregates in biosensor cells. ( a ) HEK.A53T-C/Y cells were treated with detergent-insoluble brain fractions and imaged under conditions in which excitation was applied at CFP and emission was detected at YFP wavelengths, enabling FRET-based visualization of intracellular aggregate formation. The scale bar represents 50 µm. ( b ) Quantification of intracellular aggregates in PD and control (C) samples. ( c ) Quantification of intracellular aggregates across controls, EOPD and LOPD. ( d ) Within LOPD subgroups, comparison of aggregate burden in FP-LO and SP-LO patients. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Data are shown as violin plots normalized to the negative control condition (lipofectamine-treated cells set to 1). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test for group comparisons and unpaired two-tailed Student’s t-tests for pairwise comparisons

Article Snippet: The reaction mixture contained 1X phosphate-buffered saline (KD Medical, PBS 10X, pH 8.0), 0.0006% sodium dodecyl sulfate (SDS), 10 μM Thioflavin T (ThT), and 0.1 mg/mL recombinant human aSyn monomer (Proteos, RP-010).

Techniques: Control, Comparison, Negative Control, Two Tailed Test

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Proteinase K (PK) digestion of seed amplification assay products identified differential proteolytic resistance profiles between controls (C) and PD patients. ( a ) Undigested product is shown on a Silver Stain at the expected band size for aSyn monomer (~ 15 kDa). PK digestion broke this product down over time. ( b ) PD patient products were more resistant than controls at all time points. ( c ) Although patient groups remained more resistant than controls, no differences within patient sub-groups were found in relation to disease onset. ( d ) Between late-onset patients, a trend toward higher resistance was observed in FP-LO patients. Data represent mean ± SD. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons and the Mann–Whitney U test for pairwise comparisons

Article Snippet: The reaction mixture contained 1X phosphate-buffered saline (KD Medical, PBS 10X, pH 8.0), 0.0006% sodium dodecyl sulfate (SDS), 10 μM Thioflavin T (ThT), and 0.1 mg/mL recombinant human aSyn monomer (Proteos, RP-010).

Techniques: Amplification, Silver Staining, Comparison, MANN-WHITNEY

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Gross pathology of the cingulate gyrus and correlations with Lewy body (LB) burden and biochemical measures. Representative coronal brain sections of the cingulate gyrus showing the cingulate cortex (white arrowheads) in an exceptional case ( a ), a typical case ( b ), and a non-LB control ( c ). Gross examination shows cortical atrophy and pallor of the cingulate region in PD cases compared to controls. ( d and e ) Immunohistochemistry with anti-NACP antibody in the cingulate cortex; (a) and (d) are from the same patient, and (b) and (e) are from the same patient. Inset in (d) shows an LB at higher magnification. Scale bars: 5 mm (a–c); 100 μm (d and e); 20 μm (inset). ( f ) Quantitative analysis of LB counts across PD patients did not reveal significant differences between clinical subgroups, although substantial variability was observed within each group. ( g ) Spearman correlation analyses show a positive association between LB counts and integrated FRET density, pSer129-aSyn, and total aSyn levels in PD cases

Article Snippet: The reaction mixture contained 1X phosphate-buffered saline (KD Medical, PBS 10X, pH 8.0), 0.0006% sodium dodecyl sulfate (SDS), 10 μM Thioflavin T (ThT), and 0.1 mg/mL recombinant human aSyn monomer (Proteos, RP-010).

Techniques: Control, Immunohistochemistry

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Measurement of total and pSer129-aSyn levels in detergent-insoluble brain fractions by AlphaLISA. ( a and b ) total and pSer129-aSyn levels in PD and control (C) groups. ( c and d ) total and pSer129-aSyn levels across controls, EOPD and LOPD. ( e and f ) Comparison of total and pSer129-aSyn levels in FP-LO and SP-LO. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons (c and d) and the Mann–Whitney U test for pairwise comparisons (a, b, c and e), and Student’s t-test (f)

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Control, Comparison, MANN-WHITNEY

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: aSyn seeding bioactivity measured by FRET FC assay. ( a ) Comparison of aSyn seeding activity in PD and control (C) groups. ( b ) Comparison of aSyn seeding activity in EOPD and LOPD disease subgroups. ( c ) Comparison of aSyn seeding activity in FP-LO and SP-LO subgroups. ( d ) Correlation between pSer129-aSyn levels and FRET seeding bioactivity in PD cases. ( e ) Cell viability analysis following treatment with detergent-insoluble brain fractions. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons and the Mann–Whitney U test for pairwise comparisons. For the correlation analysis, a two-tailed Spearman’s rank correlation test was used, and r and p values are indicated on the plot

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Comparison, Activity Assay, Control, MANN-WHITNEY, Two Tailed Test

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Gross pathology of the cingulate gyrus and correlations with Lewy body (LB) burden and biochemical measures. Representative coronal brain sections of the cingulate gyrus showing the cingulate cortex (white arrowheads) in an exceptional case ( a ), a typical case ( b ), and a non-LB control ( c ). Gross examination shows cortical atrophy and pallor of the cingulate region in PD cases compared to controls. ( d and e ) Immunohistochemistry with anti-NACP antibody in the cingulate cortex; (a) and (d) are from the same patient, and (b) and (e) are from the same patient. Inset in (d) shows an LB at higher magnification. Scale bars: 5 mm (a–c); 100 μm (d and e); 20 μm (inset). ( f ) Quantitative analysis of LB counts across PD patients did not reveal significant differences between clinical subgroups, although substantial variability was observed within each group. ( g ) Spearman correlation analyses show a positive association between LB counts and integrated FRET density, pSer129-aSyn, and total aSyn levels in PD cases

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Control, Immunohistochemistry

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Measurement of total and pSer129-aSyn levels in detergent-insoluble brain fractions by AlphaLISA. ( a and b ) total and pSer129-aSyn levels in PD and control (C) groups. ( c and d ) total and pSer129-aSyn levels across controls, EOPD and LOPD. ( e and f ) Comparison of total and pSer129-aSyn levels in FP-LO and SP-LO. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons (c and d) and the Mann–Whitney U test for pairwise comparisons (a, b, c and e), and Student’s t-test (f)

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Control, Comparison, MANN-WHITNEY

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: aSyn seeding bioactivity measured by FRET FC assay. ( a ) Comparison of aSyn seeding activity in PD and control (C) groups. ( b ) Comparison of aSyn seeding activity in EOPD and LOPD disease subgroups. ( c ) Comparison of aSyn seeding activity in FP-LO and SP-LO subgroups. ( d ) Correlation between pSer129-aSyn levels and FRET seeding bioactivity in PD cases. ( e ) Cell viability analysis following treatment with detergent-insoluble brain fractions. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons and the Mann–Whitney U test for pairwise comparisons. For the correlation analysis, a two-tailed Spearman’s rank correlation test was used, and r and p values are indicated on the plot

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Comparison, Activity Assay, Control, MANN-WHITNEY, Two Tailed Test

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Intracellular aSyn aggregates in biosensor cells. ( a ) HEK.A53T-C/Y cells were treated with detergent-insoluble brain fractions and imaged under conditions in which excitation was applied at CFP and emission was detected at YFP wavelengths, enabling FRET-based visualization of intracellular aggregate formation. The scale bar represents 50 µm. ( b ) Quantification of intracellular aggregates in PD and control (C) samples. ( c ) Quantification of intracellular aggregates across controls, EOPD and LOPD. ( d ) Within LOPD subgroups, comparison of aggregate burden in FP-LO and SP-LO patients. Data represent mean ± SD. Each dot represents the average value for an individual case. Violin plots show the distribution of the data, with the median indicated by the central line and quartiles by the dashed lines. Data are shown as violin plots normalized to the negative control condition (lipofectamine-treated cells set to 1). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test for group comparisons and unpaired two-tailed Student’s t-tests for pairwise comparisons

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Control, Comparison, Negative Control, Two Tailed Test

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Proteinase K (PK) digestion of seed amplification assay products identified differential proteolytic resistance profiles between controls (C) and PD patients. ( a ) Undigested product is shown on a Silver Stain at the expected band size for aSyn monomer (~ 15 kDa). PK digestion broke this product down over time. ( b ) PD patient products were more resistant than controls at all time points. ( c ) Although patient groups remained more resistant than controls, no differences within patient sub-groups were found in relation to disease onset. ( d ) Between late-onset patients, a trend toward higher resistance was observed in FP-LO patients. Data represent mean ± SD. Statistical significance was determined using Kruskal–Wallis test followed by Dunn’s multiple comparison test for group comparisons and the Mann–Whitney U test for pairwise comparisons

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Amplification, Silver Staining, Comparison, MANN-WHITNEY

Journal: Acta Neuropathologica

Article Title: Molecular profiling of alpha-synuclein pathology and seeding activity in Parkinson’s disease

doi: 10.1007/s00401-026-03026-1

Figure Lengend Snippet: Gross pathology of the cingulate gyrus and correlations with Lewy body (LB) burden and biochemical measures. Representative coronal brain sections of the cingulate gyrus showing the cingulate cortex (white arrowheads) in an exceptional case ( a ), a typical case ( b ), and a non-LB control ( c ). Gross examination shows cortical atrophy and pallor of the cingulate region in PD cases compared to controls. ( d and e ) Immunohistochemistry with anti-NACP antibody in the cingulate cortex; (a) and (d) are from the same patient, and (b) and (e) are from the same patient. Inset in (d) shows an LB at higher magnification. Scale bars: 5 mm (a–c); 100 μm (d and e); 20 μm (inset). ( f ) Quantitative analysis of LB counts across PD patients did not reveal significant differences between clinical subgroups, although substantial variability was observed within each group. ( g ) Spearman correlation analyses show a positive association between LB counts and integrated FRET density, pSer129-aSyn, and total aSyn levels in PD cases

Article Snippet: For AlphaLISA, these fractions were diluted 2.5-fold with the kit-provided lysis buffer and incubated on ice for 20 min. For quantitative measurements, recombinant human aSyn (Proteos, RP-003) and recombinant human pSer129-aSyn (Proteos, RP-004) proteins were used to generate standard curves for total and pSer129-aSyn quantification, respectively.

Techniques: Control, Immunohistochemistry