1. Organ-specific Neurodegeneration in Triple A syndrome-related Achalasia 
Vincent Zimmer, MD, Jean-Marie Vanderwinden, MD, PhD, Anna Zimmer, MD, Daniel Ostertag, MD, Matthias Strittmatter, MD, Katrin Koehler, MD, Angela Huebner, MD, Frank Lammert, MD 
The American Journal of Medicine 
Volume 128, Issue 9, Pages e9-e12 (September 2015)
DOI: /j.amjmed
Copyright © 2015 Elsevier Inc. Terms and Conditions

2. Figure 1
Molecular and cellular studies. (A) Sequencing of the 16 exons and all exon-intron boundaries of the AAAS gene was performed in the index patient II:5 first, revealing the compound heterozygous mutations c.355C>T (p.Arg119*) and c.903A>C (p.Lys301Asn) on exons 4 and 9, respectively. Genetic studies in the index patient's parents confirmed each of them to carry one AAAS gene mutation. Further genetic testing in the sibling generation indicated an older brother to be also affected by the mutational status, which was confirmed subsequently by clinical assessment (first diagnosis of early-stage achalasia, alacrima without adrenal insufficiency). Two heterozygous siblings were free of triple A syndrome-compatible clinical symptoms. (B) Immunofluorescence analysis of ALADIN (red) in cultured fibroblast cells from the index patient II:5 and a healthy control. Nuclei were stained with DAPI (blue). Cells with the p.Arg119*/p.Lys301Asn mutation show a localization of ALADIN at the nuclear pore complex comparable with control cells.
The American Journal of Medicine  , e9-e12DOI: ( /j.amjmed )
Copyright © 2015 Elsevier Inc. Terms and Conditions

3. Figure 2
Dedicated immunofluorescence analysis of the esophagocardiac neuromuscular interface. (A) PDG9.5 (pan-neuronal marker) staining only scarce nerve strands presumably of extrinsic origin in the intermuscular plane but neither detection of enteric nervous system ganglia nor thin nerve fibers in the muscle layers. In addition to severe reduction in PGP9.5 immunoreactivity, complete absence of nNOS staining likewise suggests a severe alteration of the neural component (not shown) (B) SMA (smooth muscle marker) revealing an inconspicuous distribution of immunoreactivity in muscle layers and blood. (C) SK3 (fibroblast-like cells marker) immunostaining indicates abundant signals in muscle layers as expected. (D) KIT (interstitial cells of Cajal [ICC] marker) staining with identification of typical spindle-shaped ICCs and round mast cells (strongest KIT positivity).
The American Journal of Medicine  , e9-e12DOI: ( /j.amjmed )
Copyright © 2015 Elsevier Inc. Terms and Conditions