J. Lipid Res.
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A more recent version of this article appeared on May 1, 2005

Papers In Press, published online ahead of print March 1, 2005
J. Lipid Res., doi:10.1194/jlr.M400458-JLR200
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Submitted on November 17, 2004
Revised on February 16, 2005
Accepted on February 18, 2005

Biliary cholesterol crystallization characterized by single crystal cryogenic-electron diffraction

Daphne Weihs, Judith Schmidt, Ilana Goldiner, Dganit Danino, Moshe Rubin, Yeshayahu Talmon, and Fred M. Konikoff

Department of Chemical Engineering, Technion-Israel institute of Technology, Haifa 32000

Corresponding Author: ishi{at}techunix.technion.ac.il

Cholesterol crystals are the building blocks of cholesterol gallstones. The exact structure of early forming crystals is still controversial. We combined cryogenic-temperature transmission electron microscopy with cryogenic-temperature electron diffraction to sequentially study crystal development and structure in nucleating model and native gallbladder biles. The growth and long-term stability of classic cholesterol monohydrate crystals in native and model biles was determined. In solutions of model bile with low phospholipid to cholesterol ratio electron diffraction provided direct proof of a novel transient polymorph. It had an elongated habit and unit-cell parameters differing from classic, triclinic cholesterol monohydrate. This crystal is exactly the monoclinic cholesterol monohydrate phase described by Solomonov and coworkers (Biophysical J., in press) in cholesterol monolayers compressed on the air-water interface. We observed no evidence of anhydrous cholesterol crystallization in any of the biles studied. In conclusion, classic cholesterol monohydrate is the predominant and stable form in native and model biles. However, under certain (low phospholipid) conditions, transient, intermediate polymorphs may form. These findings, documenting single crystal analysis in bulk solution, provide an experimental approach to investigate factors influencing biliary cholesterol crystal nucleation and growth as well as other processes of nucleation and crystallization in liquid systems.


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