Raw data and summary of 162 B6SJL-TgN-(SOD1-G93A)1Gur autosomal hemizygous female (F) and male (M) mice meeting the additional endpoint criteria prior to reaching CS 4.

<p>20%CS2, weight loss ≥20% <i>vs.</i> body weight immediately prior to a clinical score of 2; 20%Peak, weight loss ≥20% <i>vs.</i> peak body weight; BC<2, body condition score <2.</p><p>*Earliest age (d) of 20%CS2, 20%Peak and BC<2 was used to calculate mean age. Data for Mean Age (d) are presented as means ± SD.</p

Probability of survival for CS 4, CS 4+ and CS 5.

<p>Probability of survival for the 3 different endpoints (CS 4, black line; CS 4+, blue line; CS 5, red line). For all logrank tests, CS 4 was used as the reference when comparing CS 4 vs. CS 4+ and CS 4 vs. CS 5, whereas CS 4+ was used as the reference when comparing CS 4+ vs. CS 5. The rate of reaching endpoint is significantly different (P = 0.021) between CS 4 (clinical score of 4 = functional paralysis of both hindlimbs), CS 4+ [CS 4 plus the earliest of a) weight loss ≥20% <i>vs.</i> body weight immediately prior to a clinical score of 2, b) weight loss ≥20% <i>vs.</i> peak body weight, c) body condition score <2, or d) a righting reflex >20 s (CS 5)], and CS 5 (clinical score of 5 = CS 4 and righting reflex >20 s). Mice reached CS 4 at a rate of 34% faster vs. CS 5 (HR = 1.34; 95% CI 1.10, 1.74; P = 0.006) and 24% faster vs. CS 4+ (HR = 1.24; 95% CI 1.00, 1.59; P = 0.046). Mice reached CS 4+ at a non-significant rate of 9% faster vs. CS 5 (HR = 1.09; 95% CI 0.88, 1.38; P = 0.410).</p

Correlation between CS 4 and CS 4+ and the Bland-Altman plot for CS 4 and CS 4+.

<p>(A) Correlation between CS 4 (clinical score of 4 = functional paralysis of both hindlimbs) and CS 4+ [CS 4 plus the earliest of a) weight loss ≥20% <i>vs.</i> body weight immediately prior to a clinical score of 2, b) weight loss ≥20% <i>vs.</i> peak body weight, c) body condition score <2, or d) a righting reflex >20 s (CS 5)]. There was a strong positive relationship between CS 4 and CS 4+ (r = 0.96, slope = 0.92, P<0.001). CS 4+ (d) = (12.79±2.56)+[(0.92±0.02)×(CS 4 in d)], mean ± SEM. Dashed line indicates line of identity. (B) A Bland-Altman plot comparing CS 4 to CS 4+. Mean bias ± SD = 2.2±2.3%, lower limit = −2.4%, upper limit = 6.7%.</p

Raw data and summary of endpoint criteria for 162 B6SJL-TgN-(SOD1-G93A)1Gur autosomal hemizygous female (F) and male (M) mice.

<p>CS 4, clinical score of 4 = functional paralysis of both hindlimbs; 20%CS2, weight loss ≥20% <i>vs.</i> body weight immediately prior to a clinical score of 2; 20%Peak, weight loss ≥20% <i>vs.</i> peak body weight; BC<2, body condition score <2; CS 5, clinical score of 5 = CS 4 plus a righting reflex >20 s; CS4+, clinical score of 4+ = CS 4 in addition to the earliest of 20%CS2, 20%Peak, BC<2 or a righting reflex of >20 s.</p><p>*Significantly different from each other (P<0.001). Data for Mean Age (d) are presented as means ± SD.</p

Correlation between CS 4 and CS 5 and the Bland-Altman plot for CS 4 vs. CS 5.

<p>(A) Correlation between CS 4 (clinical score of 4 = functional paralysis of both hindlimbs) and CS 5 (clinical score of 5 = CS 4 plus a righting reflex >20 s). There was a strong positive relationship between CS 4 and CS 5 (r = 0.95, slope = 0.91, P<0.001). CS 5 (d) = (14.80±2.83)+[(0.91±0.02)×(CS 4 in d)], mean ± SEM. Dashed line indicates line of identity. (B) A Bland-Altman plot comparing CS 4 to CS 5. Mean bias ± SD = 3.0±2.5%, lower limit = −2.0%, upper limit = 7.9%.</p

Correlation between CS 5 and CS 4+ and the Bland-Altman plot for CS 5 and CS 4+.

<p>(A) Correlation between CS 5 (clinical score of 5 = CS 4 and righting reflex >20 s) and CS 4+ [CS 4 plus the earliest of a) weight loss ≥20% <i>vs.</i> body weight immediately prior to a clinical score of 2, b) weight loss ≥20% loss <i>vs.</i> peak body weight, c) body condition score <2, or d) a righting reflex >20 s (CS 5)]. There was a strong positive relationship between CS 5 and CS 4+ (r = 0.98, slope = 0.98, P<0.001). CS5 (d) = (3.93±2.15)+[(0.98±0.02)×(CS 4+ in d)], mean ± SEM. Dashed line indicates line of identity. (B) A Bland-Altman plot comparing CS 5 to CS 4+. Mean bias ± SD = 0.8±1.7%, lower limit = −2.5%, upper limit = 4.1%.</p

Neurotrophic factor in DEF vs. AI G93A mice.

<p>GDNF protein content (A and B) (arbitrary units; AU) in spinal cord of 42 G93A mice: 23 adequate vitamin D₃ intake (AI; 1 IU D₃/g feed; 12 M, 11 F) and 19 deficient vitamin D₃ intake (DEF; 0.025 IU D₃/g feed; 10 M, 9 F). There was no significant difference in GDNF protein content between the diets or between the sexes. Data presented as means ± SEM.</p

Antioxidant enzymes in DEF vs. AI G93A mice.

<p>SOD2 (A and B), catalase (C and D) and GPx1 (E and F) protein content (arbitrary units; AU) in spinal cord of 42 G93A mice: 23 adequate vitamin D₃ intake (AI; 1 IU D₃/g feed; 12 M, 11 F) and 19 deficient vitamin D₃ intake (DEF; 0.025 IU D₃/g feed; 10 M, 9 F). <u><i>SOD2 (A and B)</i>:</u> DEF males had 18% lower SOD2 protein content vs. AI males (P = 0.034). DEF males had 27% lower SOD2 protein content vs. DEF females (P = 0.004). <u><i>Catalase (C and D)</i>:</u> There was no significant difference in catalase protein content between the diets or between the sexes. <u><i>GPx1 (E and F)</i>:</u> DEF mice had 12% higher GPx1 protein content vs. AI (P = 0.057). DEF females had 29% higher GPx1 protein content vs. AI females (P = 0.001). AI males had 10% higher GPx1 protein content vs. AI females (P = 0.054). DEF males had 17% lower GPx1 protein content vs. DEF females (P = 0.070). Data presented as means ± SEM.</p

Spinal cord weight between the diets and sexes.

<p>Data are means ± SEM. AI, adequate intake; DEF, deficient vitamin D. AI Males, n = 12; AI Females, n = 11. DEF Males, n = 10; DEF Females, n = 9</p><p>Spinal cord weight between the diets and sexes.</p

Body weight-adjusted spinal cord weights.

<p>Body weight-adjusted spinal cord weight (mg/g b.wt.) of 42 G93A mice: 23 adequate vitamin D₃ intake (AI; 1 IU D₃/g feed; 12 M, 11 F) and 19 deficient vitamin D₃ intake (DEF; 0.025 IU D₃/g feed; 10 M, 9 F). <u><i>Between the diets (A and B)</i>:</u> There was no significant difference in body weight-adjusted spinal cord weights between the diets. <u><i>Between the sexes (C and D)</i>:</u> AI males had 33% lighter body weight-adjusted spinal cord weight vs. AI females (P = 0.001), and DEF males had 27% lighter body weight-adjusted spinal cord weight vs. DEF females (P = 0.005). Data presented as means ± SEM.</p