1. Volume 130, Issue 3, Pages 893-901 (March 2006)
Mitochondrial Neurogastrointestinal Encephalomyopathy: Evidence of Mitochondrial DNA Depletion in the Small Intestine 
Carla Giordano, Mariangela Sebastiani, Giuseppe Plazzi, Claudia Travaglini, Patrizio Sale, Marcello Pinti, Andrea Tancredi, Rocco Liguori, Pasquale Montagna, Marzio Bellan, Maria Lucia Valentino, Andrea Cossarizza, Michio Hirano, Giulia d’Amati, Valerio Carelli 
Gastroenterology 
Volume 130, Issue 3, Pages (March 2006)
DOI: /j.gastro
Copyright © 2006 American Gastroenterological Association Terms and Conditions

2. Figure 1
Morphology of small intestinal wall of a control subject (A) and of the MNGIE patient (B–E). (A) Masson Trichrome stain, 2.5×. The normal structure of small intestinal wall shows an internal circular layer (IL) of muscularis propria separated by the external longitudinal layer (EL) with interposed myenteric plexus (asterisk). (B) Masson Trichrome stain, 2.5×. In the MNGIE patient, the internal muscle layer (IL) of muscularis propia is preserved; by contrast, the external layer is atrophic with only a few scattered islands of muscle fibers (arrows) left within the fibrous tissue. The myenteric plexus (asterisk) appears well preserved. Note that the apparent difference in thickness of the entire section compared with the control is because of the slightly different plane of cut. (C) H&E, 20×. Smooth-muscle fibers in the atrophic external layer (EL) show extensive cytoplasmic microvacuolation (arrows) and picnotic nuclei. Nervous tissue is normal (asterisk). (D) S-100, 10×; insert, synaptophysin 40×. The myenteric plexus appears well preserved and shows normal distribution. A normal looking ganglion cell is shown in the insert. (E) Ultrastructural features of smooth-muscle cells in the small intestine of the MNGIE patient. Smooth-muscle cells are recognized by the presence of “focal adhesions” (square boxes). The cell cytoplasm has a vacuolated appearance being filled by numerous empty profiles of swollen mitochondria (asterisks) and lipid droplets (arrows). Note the shrunken nucleus with condensed chromatin.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Terms and Conditions

3. Figure 2
Evaluation of mtDNA copy number in GI tissues homogenates. (A) Interaction plot of mtDNA copies/cells (log scale) means versus GI tissue-segment homogenates for the proband (red line) and 10 controls (black lines). The ileum, in the 10 controls subjects, has a lower amount of mtDNAs relative to esophagus, stomach, and colon. The MNGIE patient shows a striking depletion limited to the small intestine. (B) Residual plot: for each point, the x-value is the estimate of mtDNA copy number obtained with the mixed-effect model (see Statistical Analysis in the Methods section), and the y-value is the standardized difference (residual) between the estimate and the observed value. High standardized residuals, as we observed for the MNGIE patient ileum (red point within large black circle), may be considered outliers, a distinct cluster because of the disease.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Terms and Conditions

4. Figure 3
LCM and evaluation of mtDNA copy number in different tissues from the small intestine wall. (A) Small intestine wall of MNGIE patient (H&E, 20×) before (right) and after (middle) microdissection of cells from the internal (IL) and external (EL) layers of muscularis propria and the autonomic nervous tissue of the myenteric plexus (asterisk). Microdissected tissues are shown (left). (B) Interaction plot of mtDNA copies/cells means versus microdissected tissues of small intestine for the proband (red line) and 5 controls (black lines). In controls, the mtDNA copy number is lower in the external layer of muscularis propria compared with the internal one (internal/external layer ratio 2:1). The MNGIE patient shows a profound depletion limited to the external layer of muscularis propria. (C) Residual plot: for each point, the x-value is the estimate of mtDNA copy number obtained with the mixed effect model (see Statistical Analysis in the Methods section) and the y-value is the standardized difference (residual) between the estimate and the observed value. Red point represents the patient.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Terms and Conditions