title: Intramitochondrial crystalline inclusions in nonalcoholic steatohepatitis.
abstract: Mitochondrial dysfunction is an important element in the pathogenesis of nonalcoholic steatohepatitis (NASH). Intramitochondrial crystals (IMCs) are a well-documented morphological abnormality seen on transmission electron microscopy in this disease. It has been suggested that IMCs consist of phospholipids, but their exact composition remain uncertain many years after their discovery. Micellar phase transitions of phospholipid bilayers is a well-known but little-studied phenomenon in living systems. Its presence in the mitochondria of NASH would offer significant insight into the disease with possible therapeutic implications. We postulated that intramitochondrial disturbances in NASH are sufficient to produce such transitions and that their detection in fresh biopsies would therefore be a dynamic process. To test this, we performed a blinded, prospective analysis of fresh liver biopsy samples immediately fixed under different conditions. Quantitative transmission electron microscopy morphometry, performed by systematically counting total mitochondria and IMCs within areas of uniform dimension, showed a stepwise decline in IMCs with cooler fixation temperature in each subject studied. Randomization testing (Monte Carlo resampling) confirmed that the detection of IMCs was strongly dependent on fixation temperature (P < 0.0001).
  
  
  CONCLUSIONS
  These results indicate that the intramitochondrial crystals characteristic of NASH are highly dynamic and unstable structures. The findings offer the strongest support yet for their origin in micellar phase transitions. We speculate that such transitions result from microenvironmental changes within the mitochondria and carry therapeutic implications, especially in regard to dietary manipulations of mitochondrial lipid composition.
authors.affiliation: Division of GI/Hepatology, Department of Internal Medicine, University of Virginia, Charlottesville, VA 22908-0708, USA. shc5c@virginia.edu
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Caldwell
authors.forename: Luiz Antonio R
authors.initials: LA
authors.lastname: de Freitas
authors.forename: Sang H
authors.initials: SH
authors.lastname: Park
authors.forename: Maria Lucia V
authors.initials: ML
authors.lastname: Moreno
authors.forename: Jan A
authors.initials: JA
authors.lastname: Redick
authors.forename: Christine A
authors.initials: CA
authors.lastname: Davis
authors.forename: Barbee J
authors.initials: BJ
authors.lastname: Sisson
authors.forename: James T
authors.initials: JT
authors.lastname: Patrie
authors.forename: Helma
authors.initials: H
authors.lastname: Cotrim
authors.forename: Curtis K
authors.initials: CK
authors.lastname: Argo
authors.forename: Abdullah
authors.initials: A
authors.lastname: Al-Osaimi
mesh_terms: D000328:Adult; D000368:Aged; D003460:Crystallization; D005234:Fatty Liver / Q000473:pathology*; D005260:Female; D006801:Humans; D008875:Middle Aged; D008928:Mitochondria / Q000473:pathology*; D011446:Prospective Studies; D016037:Single-Blind Method