IAPR Workshop 017 CV- Speaal Hardware and Industrial Applications OCT.12-14. 1988. Tokyo A SEGMENT-BASED MATCHING ALGORITHM IN TRINOCULAR VISION

ABSTRACT been proposed to improve matching accurac

Reduced venusYFP fluorescence in the presence of CHIP.

<p>Coronal sections of the SNpc were immunostained with anti-Myc (alexa-635, purple) to evaluate co-expression of CHIP with α-syn aggregates (venusYFP, green). Images revealed extensive co-expression of CHIP and α-syn aggregates demonstrating efficient viral co-transduction (A). Coronal Sections were imaged for venusYFP fluorescence and α-syn immunostaining (red) to evaluate the level of α-syn aggregates (B). Image analysis demonstrates in syn+CHIP group (+CHIP) a significant reduction in the overall venusYFP fluorescence and no change in α-syn immunostaining compared to syn-CHIP (−CHIP)(C). Detailed image analysis shows a significant 35% reduction in the number of venusYFP positive cells in the syn+CHIP group compared to syn-CHIP (D) as well as a 4-fold reduction in venusYFP fluorescence per cell compared to control (E). Representative images are displayed. Scale bars 200 µm.</p

CHIP reduces the amount of human α-syn in the striatum.

<p>Striatal homogenates were biochemically analyzed for α-syn levels. Higher molecular weight α-syn species are visible in native-PAGE (A). Band quantification revealed a 43% reduction in α-syn aggregates measured in the syn+CHIP group (+CHIP) compared to the syn-CHIP group (−CHIP) (B). Denatured samples were also run on SDS-PAGE to evaluate the total amount of α-syn. The higher band at ∼31.3 KD and lower band at ∼22.4 KD detected with anti-α-syn correspond to V1S and SV2 respectively (C). A 26% and a 69% reduction were measured in both V1S and SV2 respectively in animals expressing CHIP (D and E).</p

NeuN quantification in the presence of CHIP.

<p>Blinded stereological analysis of NeuN immunopositive cells in coronal sections across the SNpc was performed. Sections were co-stained with anti TH antibody (A., purple) and NeuN (B., red). A binary image was produced where the SNpc is delineated according to the TH immunostaining (C., blue). Comparison of percent cell loss per animal revealed discrepancies in the TH measured lesion compared to NeuN in the presence of CHIP (E), unlike animals not expressing CHIP (D). We found a 20–30% difference in lesions determined by NeuN to that determined by TH in the presence of CHIP (F). A significant 23% NeuN lesion was measured in the syn–CHIP group whereas no significant lesion was measured in the venus+CHIP group and syn+CHIP (F). Scale bars 200 µm. For the purpose of illustrating the image analysis conducted in this assay only representative images of CHIP animal are presented.</p

TH cell loss in the presence of CHIP.

<p>Unbiased blinded stereological analysis of TH immunopositive cells in coronal sections across the SNpc was performed using DAB (A). Cell loss is determined by the ratio of TH positive cells ipsilateral (right, asterisk) to contralateral (left). A 34% lesion and a 35% lesion were measured in the syn–CHIP group (n = 10, p<0.005) and syn+CHIP group (n = 7, p<0.005) respectively (B). The venus+CHIP group expressing only venusYFP and CHIP had a 44% lesion (n = 4, p<0.05). Representative images are displayed. Scale bar 500 µm.</p

Comparison of higher dose SNX-0723 and SNX-9114 effects on nigrostriatal toxicity. A–D

<p>) Photos show low power images of injected SN (right) and contralateral uninjected side (left) for each drug treatment group. Black (medial) and white (lateral) squares indicate regions of interest for higher magnification photos shown. There is modest cell loss on the side of the lesion for all treatments. <b>E</b>) Graph of stereological counts (mean ±SEM) of TH-positive cells in the SN ipsilateral and contralateral to AAV-α-synuclein lesion. Numbers at base of bars indicate N for each group. Analysis of variance revealed no significant differences among treatment groups.</p

Graph of cumulative weight gain for treatment groups.

<p>SNX-0723 caused the most toxicity, weight loss, and failure to thrive at the higher dose of 10 mg/kg. All treatment groups gained weight at lower doses, including SNX-9114, but not at the rate of vehicle control (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 for comparison of vehicle and 9114 at 1.5 mg/kg; 2-way ANOVA with Bonferroni correction). (n  = 14, 14, 12, 10, and 14 for vehicle, 0723 [3mg/kg], 0723 [6–10 mg/kg], 9114 [1.5 mg/kg] and 9114 [3 mg/kg], respectively).</p

Hsp70 induction in brain.

<p><b>A</b>) Graph of hsp70 ELISA data from cortical tissue lysates after treatment of rats with novel small molecule Hsp90 inhibitors. Tissue was harvested at 1–6 days post treatment and shows sustained hsp70 induction at ≥72 hrs for both SNX-0723 and SNX-9114. <b>B</b>) Western blot of striatal tissue homogenates from rats injected with WT α-synuclein and treated with SNX9114 (n = 5) or vehicle (n = 9), immunoblotted for hsp70 and actin as loading control. <b>C</b>) Densitometry shows significant (p = 0.037) striatal Hsp70 induction following treatment with SNX-9114.</p

Illustration of drug effects on nigrostriatal terminal density for SNX-9114, 3 mg/kg dose.

<p>The representative photos show the distribution of TH+ terminals in the striatum contralateral and ipsilateral to AAV-α-synuclein injection in panels <b>A</b> and <b>B,</b> respectively. <b>C)</b> α-Synuclein-positive nigrostriatal terminals ipsilateral to AAV injection in same case. <b>D</b>) Photo of TH+ terminals in striatum from animal treated with lower dose SNX-9114, 1.5 mg/kg. *Marks region of TH terminal loss due to α-synuclein toxicity.</p

Study paradigm and structure of compounds.

<p><b>A</b>) Illustrates the study paradigm and timecourse for drug testing in animals. <b>B</b>) Shows the structures of parent and the two derivative SNX compounds used in experiments.</p