Supporting Research These works are/were partially supported by the National Institutes of Health: National Institute on Aging SBIR grant No. AG037214. The views, opinions and/or findings contained in this work are those of Actuated Medical, Inc. and should not be construed as an official government position, policy or decision unless so designated by other documentation. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Reduced Plasma Corticosterone for Serial Blood Collection GentleSharp reduces the average plasma corticosterone concentration obtained from rat tail vein samples during serial blood collections. Results With GentleSharp ON during needle puncture (orange bar), the average plasma corticosterone concentration is reduced compared to GentleSharp OFF during puncture (gray bar) on week 2 (49.2% reduction) and 3 (65.2% reduction) of blood collections. Repeated Measures Analysis of Variance, statistical significance of mean difference within week of collection: * p < 0.05 Corticosterone (ng/mL) Summary 1000 * p < 0.05 OFF ON * 800 * 600 400 200 0 WEEK 1 Reduction: 9.7% WEEK 2 49.2% WEEK 3 65.2% Experimental Design > Sprague Dawley Rats (n =19). > 9 total collection trials per subject - 3 sampling days (1 week apart) with 3 collections / day spaced 60 min apart. > Week 1 - ON group: n = 19 trials with 10 rats; OFF group: n = 21 trials with 9 rats; OFF vs ON groups, compared within week; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Reduced Animal Movement and Vocalization for Serial Blood Collection GentleSharp reduces the average animal vocalization (VOCAL) and movement (MOVE) during serial blood sampling from rat tail veins. Results With GentleSharp ON during needle puncture (orange bars), the average animal movement (MOVE) and vocalization (VOCAL) is reduced compared to GentleSharp OFF during puncture (gray bars) on weeks 2 and 3 of blood collections. Student’s t-test, statistical significance of mean difference within week of collection: * p < 0.05 ** p < 0.005 Average Rating (1 - 5 Scale) Summary 3.0 * p < 0.05 ** p < 0.005 ** ** 2.5 OFF ON * * 2.0 1.5 1.0 0.5 0.0 0 VOCAL MOVE WEEK 1 VOCAL MOVE WEEK 2 VOCAL MOVE WEEK 3 Experimental Design > Sprague Dawley Rats (n = 19). > 9 total collection trials per subject - 3 sampling days (1 week apart) with 3 collections / day spaced 60 min apart. > Videos of collections viewed and graded (Likert Scale, 1 - 5) by blinded reviewers. > ON group: n = 30 trials with 10 rats; OFF group: n = 27 trials with 9 rats; OFF vs ON groups, compared within week; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Reduced Peak Insertion Force During Rat Tail Penetration Summary GentleSharp reduces the peak insertion force required for needle insertions into cadaver rat tails. 16 12 8 4 0 (cm) Difference = 67.5% ** p < 0.001 1.4 1.2 1.4 1.2 Peak Force (n) With GentleSharp ON during needle puncture (orange bar), the average peak insertion force for needle insertions into rat cadaver tails was reduced by 67.5% compared to GentleSharp OFF during puncture (gray bar). Peak Force (n) Results 1.8 1.6 1.0 0.8 0.6 0.4 0.2 0.0 OFF ON 0.08 0.06 ** 0.04 0.02 1 Student’s t-test, statistical significance of mean difference: ** p < 0.001 1.0 2 3 Tail Section > INDIVIDUAL TRIALS < 4 0.0 OFF ON > AVERAGE MEANS OF ALL TRIALS < Experimental Design > Cadaver Sprague Dawley Rat Tails (n = 6) at 3 separate locations along length of tail as shown. > ON n = 18; OFF n = 18; Error bars = standard deviation. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Decreased Needle Insertions for Serial Blood Collection Summary Results With GentleSharp ON during needle puncture (orange bar), the average number of needle insertions required to obtain a successful sample was significantly reduced compared to the GentleSharp OFF during puncture (gray bar). Repeated Measures Analysis of Variance, statistical significance of mean difference: ** p < 0.01 ** p < 0.01 2.5 Needle Insertions GentleSharp reduces the average number of needle insertions required to obtain a successful blood sample from mice tail arteries during serial blood collections. ** 2.0 1.5 1.0 0.5 0.0 OFF ON Experimental Design > C57BL/6J mice (n = 47), starting at ~11 wks of age. > 9 total collection trials per subject - 3 sampling days (1 week apart) with 3 collections / day spaced 30 min apart; each trial limited to 3 insertion attempts. > ON group: n = 216 trials with 24 mice; OFF group: n = 207 trials with 23 mice; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Increased Collection Success for Serial Blood Collection 90 GentleSharp increases the average success rate for serial blood collection from mice tail arteries. 85 Results For the group with GentleSharp ON (orange bar) during needle puncture, there was a significantly higher success rate compared to the group punctured with GentleSharp OFF (gray bar). Success is defined as obtaining ≥ 0.01 g total blood mass (~10 µl blood) per trial; individual success rate based on 9 collection trials per animal (e.g. 9:9 = 100%). Success (%) Summary ** p < 0.005 ** 80 75 70 65 60 55 50 OFF ON Average Success Rate 64.7 ± 3.4% 80.1 ± 3.5% Proportion with 100% Success 0/23 = 0% 6/24 = 25% Experimental Design Student’s t-test, statistical significance of mean difference: ** p < 0.005 > C57BL/6J mice (n = 47), starting at ~11 wks of age. > 9 total collection trials per subject - 3 sampling days (1 week apart) with 3 collections / day spaced 30 min apart; each trial limited to 3 insertion attempts. > ON group: n = 24 mice; OFF group: n = 23 mice; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Decreased Needle Insertions for Serial Blood Collection - Repeated Sampling On Same Day Summary Results With GentleSharp ON during needle puncture (orange bars), the average number of needle insertions required to successfully obtain sample was significantly reduced compared to the GentleSharp OFF during puncture (gray bars). Repeated Measures Analysis of Variance, statistical significance of mean difference within collection: * p < 0.05 OFF ON * p < 0.05 Needle Insertions GentleSharp reduces the average number of needle insertions required for successful blood collection from mice tail arteries during serial blood collections within same collection day. 2.6 2.4 2.2 2 1.8 1.6 * 1.4 1.2 1 Collection 1 Collection 2 Collection 3 Experimental Design > C57BL/6J mice (n = 47), starting at ~11 wks of age. > 9 total collection trials per subject - sampling days (1 week apart) with 3 collections / day spaced 30 min apart; each trial limited to 3 insertion attempts. > ON group: n = 72 trials with 24 mice; OFF group: n = 69 trials with 23 mice; OFF vs ON groups, compared within each collection separately; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip Summary Results Novice collectors exhibited greater differences in success rates between collections completed with GentleSharp ON (orange bars) vs OFF (gray bars) during needle puncture compared to Trained collectors. Where procedures performed with GentleSharp ON, the gap in success rate between Novice and Trained was essentially eliminated. Experiment 2 Experiment 1 OFF ON OFF ON 100 Success (%) GentleSharp increases the average success rate for novice collectors to successfully obtain blood samples from mice tail arteries, enabling them to achieve the performance of a trained collector quicker. 100 Success (%) A More Humane Blood Sampling System Increased Collection Success for Novice Collectors 80 60 40 20 0 Novice Trained 80 60 40 20 0 Novice Trained Month 1 (M1) Novice Trained OFF 5/11 10/12 M1 OFF ON 12/12 11/12 Novice Trained Month 2 (M2) Novice Trained 3/7 101/113 M1 ON 8/8 107/113 M2 OFF 10/15 117/129 M2 ON 16/16 122/129 Experimental Design > C57BL/6J mice, starting at ~11 wks of age; Target: Tail Artery. > Success is defined as obtaining sufficient blood for performance of RIA for plasma corticosterone; success rate based on successful collections / total collections attempted. Sample Sizes for Collections represented below each graph (successful trial / total trials). > Experiment 1 was completed within a single collection day. > Experiment 2 was completed over two months: M1= Month 1 totals; M2 = Month 2 cumulative totals. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System Increased Blood Mass for Serial Blood Collection Summary Results With GentleSharp ON during needle puncture (orange bar), the average blood mass was 26.7% greater compared to GentleSharp OFF during puncture (gray bar). 0.045 0.040 Blood Mass (g) GentleSharp increased the average blood mass collected per trial during serial blood collections from mice tail arteries. Difference = 26.7% * p = 0.05 * 0.035 0.030 0.025 0.020 0.015 0.010 0.005 0.000 OFF Repeated Measures Analysis of Variance, statistical significance of mean difference: * p = 0.05 ON Experimental Design > C57BL/6J mice (n = 47), starting at ~11 weeks of age. > 9 total collection trials per subject - 3 sampling days (1 week apart) with 3 collections / day spaced 30 min apart. > ON group: n = 215 trials with 24 mice; OFF group: n = 205 trials with 23 mice; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip A More Humane Blood Sampling System No Significant Change in Mice Handling Time for Serial Blood Collection Summary Results With GentleSharp ON (orange bar) during needle puncture, the mean handling time for collection trended to be reduced compared to the GentleSharp OFF during puncture (gray bar). Repeated Measures Analysis of Variance, statistical significance of mean difference: p = 0.19 105 Handling Time (s) GentleSharp does not alter the animal handling time required for blood collections during serial blood collections from mice tail arteries. 100 95 90 85 80 OFF ON Experimental Design > C57BL/6J mice (n = 47), starting at ~11 weeks of age. > 9 total collection trials per subject - 3 sampling days (1 week apart) with 3 collections / day spaced 30 min apart. > ON group: n = 212 trials with 24 mice; OFF group: n = 203 trials with 23 mice; Error bars = standard error of the mean. Learn more at GentleSharp.com CONTACT SALES +01 (814) 355-0003 x117 [email protected] ART-5022-3 Rev04 / ECO #4097 Pat. ActuatedMedical.com/ip
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