Histological evaluation of muscle biopsies from postoperative (POD) 3 to 9.

<p>Number of biopsies taken on each postoperative day (POD 3,5,7,9) in muscle according to their histological grading (Grad 0-III) based on H&E grading. The first biopsy was taken from the lateral proximal part of the thigh, the second one from the lateral distal thigh, the third one from the ventral thigh and the last one from the medial part of the thigh.</p

Results from Random Forest classification of the different rejection groups (ISO, TAC, ATC) using over the whole time course (POD 3, POD 5, POD 7, POD 9) measured inflammatory mediators in skin (A) and muscle (B) samples of rat limb transplantation models.

<p>Most important mediators for the decision trees based classification approach are evident by ranked mean decrease accuracy. Performances of the classifiers are indicated by the confusion table and the out-of-bag (OOB) error rate.</p

Similarity of sample groups and association between inflammatory mediators in rat limb transplantation models based on their profiles of mean levels in each condition.

<p>Heatmap as a result of complete linkage hierarchical clustering on log2-transformed and mean centered data. Log2-fold differences against the respective mean levels of each inflammatory mediator are color coded (red means higher inflammatory mediator levels and blue means lower inflammatory mediator levels than the mean levels of the respective inflammatory mediator according to the color scheme at the top). Dendrograms (trees) show similarity between different conditions and different inflammatory mediator profiles, respectively.</p

Appearance (Grading for skin rejection: I–IV) of each allograft on POD 1–11.

<p>*This animal died on POD 3 during the anesthesia. Bold numbers mark biopsy timepoints (skin and muscle) for each animal.</p

Results for multinomial logistic regression models in skin and muscle for classification of the rejection groups based inflammatory mediator/chemokine levels at early postoperative time points (POD 3/5) in rat transplantation models.

<p>Best prediction models for skin and muscle and their respective logistic prediction functions and classifier performance including confusion table (assessed by an 8-fold cross validation procedure) are summarized.</p

Histological evaluation of skin and muscle rejection in the early postoperative phase (POD 3 and 5).

<p>(A–C) Skin sample taken on POD 3 from an allograft without immunosuppression (A), from an isograft (B) and an allograft under TAC (C) showing no/rare inflammatory response (Grade 0 rejection). (D–F) Skin biopsies taken on POD 5 from a rejecting animal (D) displaying Grade 1 rejection, from an isograft (E) showing no/rare inflammatory response and a TAC treated allograft (F) characterized by a mild inflammatory response (Grade 0-I) in the deep dermis. (a–c) Muscle sample taken on POD 3 from an allograft without immunosuppression (a), from an isograft (b) and an allograft under TAC (c) showing no/rare inflammatory response (Grade 0 rejection). (d–f) Skin biopsies taken on POD 5 from a rejecting animal (d) displaying a mild inflammatory response (Grad 0-I rejection), from an isograft (E) and a TAC treated allograft (f) showing no/rare inflammatory response.</p

Distribution of inflammatory mediator levels at the earliest postoperative measurements (POD 3) in rat limb transplantation models.

<p>Adjusted p-values from Wilcoxon rank-sum test between the rejection group (ATC) vs. Tacrolimus treated group (TAC) for selected inflammatory mediators, which were tested to be included in a prediction model by multinomial logistic regression analysis, are presented.</p

Histological evaluation of skin biopsies from postoperative (POD) 3 to 9.

<p>Number of biopsies taken on each postoperative day (POD 3,5,7,9) in skin according to their histological grading (Grad 0-III) based on H&E staining.</p

Statistical analysis for all 14 inflammatory mediator levels in skin and in muscle at early postoperative time points (combined group of POD 3 and POD 5).

<p>Most promising inflammatory mediators for classification of rejection, based on early identification before histological manifestation of rejection is evident, are selected (<i>p_KW</i><0.05 and <i>Partition on DVs</i> >1%, symbols in bold) and subjected to multinomial logistic regression analyses. For all measured inflammatory mediators adjusted p-values from Kruskal-Wallis test (KW) of overall equality between all 3 cohorts (ISO, TAC, ATC) and Wilcoxon rank-sum test (WR) for comparison between the rejection group (ATC) and the Tacrolimus treated group (TAC) as well as partition of each inflammatory mediator on the two discriminant vectors (DVs), which maximizes the separation between the 3 rejection groups, resulting from discriminant function analysis, are given.</p

Study groups and design.

<p>* 1 mg/kg/day (for 11 days).</p