Fiber type composition, organization, and distribution are key elements in muscle functioning. These properties can be modified by intrinsic and/or extrinsic factors, such as undernutrition and injuries. Currently, there is no methodology to quantitatively analyze such modifications. On one hand, we propose a fractal approach to determine fiber type organization, using the fractal correlation method in software Fractalyse. On the other hand, we applied the kernel methodology from machine learning to build radial-basis functions for the spatial distribution of fibers (distribution functions), by dividing into square cells a two-dimensional binary image for the spatial distribution of fibers from a muscle fascicle and mounting on each cell a radial-basis function in such a way that the sum of all cell functions creates a smooth version of the fiber histogram on the cell grid. The distribution functions thus created belong in a reproducing kernel Hilbert space which permits us to regard them as vectors and measure distances and angles between them. In the present study, we analyze fiber type organization and distribution in fascicles (F2, F3, F4, and F5) of the extensor digitorum longus muscle (EDLm) from control and undernourished male rats. Fibers were classified according to the ATPase activity in slow, intermediate, and fast. Then, (x, y) coordinates of fibers were used to build binary images and distribution functions for each fiber type and both conditions. The fractal organization analysis showed that fast and intermediate fibers, from both groups, had a fractal organization within the four fascicles, i.e., the fiber assembly is distributed in clusters. We also show that chronic undernutrition altered the organization of fast fibers in the F3, although it still is considered a fractal organization. Distribution function analysis showed that each fiber type (slow, intermediate, and fast) has a unique distribution within the fascicles, in both conditions. However, chronic undernutrition modified the intra-fascicular fiber type distributions, except in the F2. Altogether, these results showed that the methodology herein proposed allows for analyzing fiber type organization and distribution modifications. On the other side, we show that chronic undernutrition alters not only the fiber type composition but also the organization and distribution, which could affect the muscle functioning, and ultimately, its behavior (e.g., locomotion).
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