ed rank test was performed for each pixel by assessing the power of the respective frequency at the observed time versus the power of the frequency at baseline. Pixels that either had a p < 0.05 (pooled response to the interdigital pinch) or p < 0.1 (single experimental group, all other stimuli) are shown. Hence, only significant changes that occurred in clusters are discussed. The cluster approach was used because of the low probability of the concentration of spurious false positives in a cluster. Therefore, the strategy was used to define a cluster size [33], i.e., either a 4 s × 2 Hz cluster for the pooled reaction to the interdigital pinch stimulus or a 4 s × 4 Hz cluster for the reaction to other stimuli. The area under the receiver operating curve (AUC) was calculated with 10k-fold bootstrapped 95% confidence intervals to track the changes in the EEG band power over time. If the 95% confidence intervals did not cross the 0.5 line, this result was considered to be significant as long as it was observed in at least at two neighbouring time points. Similar approaches have been used previously [34,35]. MATLAB R2017a (The MathWorks, Inc., Natick, MA, United States) was used for the statistical analysis. For the AUC calculation, additional functions from the measures of the effect size toolbox were used [36]. Different settings were used for the interdigital pinch stimulus and the other noxious stimuli, as a much larger number of samples was available for the interdigital pinch by pooling data from all animals, independent of the treatment groups. Beeswarm plots were generated with the MATLAB plotSpread function. 2.3.2. FOS Protein Four tissue sections per animal were included in the analysis: one section each from spinal cord segments L1, L2, L3 and S1-3. The
Area Under the Curve
Filters
ed rank test was performed for each pixel by assessing the power of the respective frequency at the observed time versus the power of the frequency at baseline. Pixels that either had a p < 0.05 (pooled response to the interdigital pinch) or p < 0.1 (single experimental group, all other stimuli) are shown. Hence, only significant changes that occurred in clusters are discussed. The cluster approach was used because of the low probability of the concentration of spurious false positives in a cluster. Therefore, the strategy was used to define a cluster size [33], i.e., either a 4 s × 2 Hz cluster for the pooled reaction to the interdigital pinch stimulus or a 4 s × 4 Hz cluster for the reaction to other stimuli. The area under the receiver operating curve (AUC) was calculated with 10k-fold bootstrapped 95% confidence intervals to track the changes in the EEG band power over time. If the 95% confidence intervals did not cross the 0.5 line, this result was considered to be significant as long as it was observed in at least at two neighbouring time points. Similar approaches have been used previously [34,35]. MATLAB R2017a (The MathWorks, Inc., Natick, MA, United States) was used for the statistical analysis. For the AUC calculation, additional functions from the measures of the effect size toolbox were used [36]. Different settings were used for the interdigital pinch stimulus and the other noxious stimuli, as a much larger number of samples was available for the interdigital pinch by pooling data from all animals, independent of the treatment groups. Beeswarm plots were generated with the MATLAB plotSpread function. 2.3.2. FOS Protein Four tissue sections per animal were included in the analysis: one section each from spinal cord segments L1, L2, L3 and S1-3. The
4). AUC values above 0.90 represent high discriminatory capacity (accuracy) of the scale [37]. The gray region represents the 95% confidence interval. https://doi.org/10.1371/journal.pone.0284218.g009 PLOS ONE | https://doi.org/10.1371/journal.pone.0284218 April 13, 2023 21 / 26 PLOS ONE Pain scale validation in pre-weaned piglets undergoing castration this painful procedure. Except for surgery, females underwent the same management and han- dling to simulate the same non-painful but potentially stressful conditions as males. The assessment of all castrated piglets 24h prior to castration served as a baseline for male piglet behavior prior to castration. The validation of this tool at the research level and by remote assessment does not automat- ically guarantee that it may be applicable for live observations. Before UPAPS is deemed a valid instrument for use in both clinical and commercial settings devoid of remote assessment, this tool will need to be validated for live observation. Lastly, pre-weaned piglets spend signifi- cant periods of their time budget sleeping. Randomly selecting video clips can be challenging if piglets are asleep. Thus, future work must take into account how to randomly select video clips to assess piglet behavior, while maximizing opportunities to view active piglets. Conclusion The UPAPS is a valid and reliable tool for remote assessment of post-castration pain in pre- weaned piglets. Further work with this tool is needed to validate and refine the scale for live observations on commercial swine farms utilizing caretakers and veterinarians responsible for animal care and management. Supporting information S1 Fig. Biplot of the principal component analysis of the UPAPS+nursing items applied to all castrated piglets (n = 39).
…AUC0–24 = area under curve from pre-treatment to 24 h post-treatment; AUC0–40 min = area under the curve from pre-treatment to 40 min post-treatment; AUC40 min–1.5…
4). AUC values above 0.90 represent high discriminatory capacity (accuracy) of the scale [37]. The gray region represents the 95% confidence interval. https://doi.org/10.1371/journal.pone.0284218.g009 PLOS ONE | https://doi.org/10.1371/journal.pone.0284218 April 13, 2023 21 / 26 PLOS ONE Pain scale validation in pre-weaned piglets undergoing castration this painful procedure. Except for surgery, females underwent the same management and han- dling to simulate the same non-painful but potentially stressful conditions as males. The assessment of all castrated piglets 24h prior to castration served as a baseline for male piglet behavior prior to castration. The validation of this tool at the research level and by remote assessment does not automat- ically guarantee that it may be applicable for live observations. Before UPAPS is deemed a valid instrument for use in both clinical and commercial settings devoid of remote assessment, this tool will need to be validated for live observation. Lastly, pre-weaned piglets spend signifi- cant periods of their time budget sleeping. Randomly selecting video clips can be challenging if piglets are asleep. Thus, future work must take into account how to randomly select video clips to assess piglet behavior, while maximizing opportunities to view active piglets. Conclusion The UPAPS is a valid and reliable tool for remote assessment of post-castration pain in pre- weaned piglets. Further work with this tool is needed to validate and refine the scale for live observations on commercial swine farms utilizing caretakers and veterinarians responsible for animal care and management. Supporting information S1 Fig. Biplot of the principal component analysis of the UPAPS+nursing items applied to all castrated piglets (n = 39).
tandard curve was generated for each assay using haptoglobin standards at 2.5, 1.25, 0.625, 0.312, and 0 mg/mL. The Page 5/20 assay procedure involved dispensing reagents…
…Peak plasma cortiso! concentrations (nmol/L) and area under the cortisol curve (AUC, nmol/L x h) during the intervals 0 to 6 h, and 6 to 24 h, 0 to 24…
…The methaemoglobin solution (0.12 mg/mL in 0.15 mol/L NaCl solution) area under the curve (AUC) from 0 to 6h also showed the same was added to all wells…
tandard curve was generated for each assay using haptoglobin standards at 2.5, 1.25, 0.625, 0.312, and 0 mg/mL. The Page 5/20 assay procedure involved dispensing reagents…
…ROC curves were then used to estimate area under the curve (AUC), sensitivity and specificity. AUC were compared statistically using DeLong test. Results Reliability Overall inter-observer reliability was “very good”, intra…