Fig. 1 Family pedigree of Patient 1 (upper) and Patient 2 (lower).
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2 Fig. 1 Family pedigree of Patient 1 (upper) and Patient 2 (lower). Fig. 2 Determination of cholesteryl ester net transfer rate from HDL to VLDL and LDL.
3 Table 1 Serum lipids and apoprotein levels in the subjects investigated Table 2 Lipid composition of lipoprotein fractions in Patient 1, Patient 2 and his family members
4 Fig. 3 Cholesteryl ester net transfer rate from HDL to VLDL and LDL in the subjects investigated. Table 3 PHLA of the subjects investigated Fig. 4 Effect of probucol on serum lipids, apo A-I and apo E levels in Patient 1.
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8 fractions:studies of humans and rabbits. Metabolism, 28:230 (1979). 4) Matsuzawa, Y., Yamashita, S., Kameda, K., Kubo, M., Tarui, S. and Hara, I.:Marked hyper-hdl2- cholesterolemia associated with premature corneal opacity-a case report. Atherosclerosis, 53:207 (1984). 5) Havel, R. J., Eder, H. A. and Bragdon, J. H.:The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum. J. Clin. Invest., 34:1345 (1955). 6) Okazaki, M., Hagiwara, N. and Hara, I.:Highperformance liquid chromatography of human serum lipoproteins;selective detection of cholinecontaining phospholipids by enzymatic reaction. J. Chromatogr., 231:13 (1982). 7) Nozaki, S., Kubo, M., Matsuzawa, Y. and Tarui, S.:Sensitive non-radioisotopic method for measuring lipoprotein lipase and hepatic triglyceride lipase in post-heparin plasma. Clin. Chem., 30: 748 (1984). 8) Fielding, P. E., Fielding, C. J. and Havel, R. J.: Cholesterol net transport, esterification, and transfer in human hyperlipidemic plasma. J. Clin. Invest., 71:449 (1983). 9) Folch, J., Lees, M. and Stanley, G. H. S.:A simple method for the isolation and purification of total lipides from animal tissues. J. Biol. Chem., 226: 497 (1957). 10) Hunninghake, D. B., Bell, C. and Olson, L.:Effect of probucol on plasma lipids and lipoproteins in type IIb hyperlipoproteinemia. Atherosclerosis, 37: 469 (1980). 11) Glueck, C. J., Gartside, P., Fallat, R. W., Sielski, J. and Steiner, P. M.:Longevity syndromes:familial hypobeta and familial hyperalpha lipoproteinemia. J. Lab. Clin. Med., 88:941 (1976). 12) Saito, F.:A pedigree of homozygous familial hyperalphalipoproteinemia. Metabolism, 33:629 (1984). 15) Barter, P. J. and Lally, J. I.:In vitro exchanges of esterified cholesterol between serum lipoprotein 16) Nichols, A. V. and Smith, L.:Effect of very-low density lipoproteins on lipid transfer in incubated serum. J. Lipid Res., 6:206 (1965). 17) Fielding, P. E. and Fielding C. J.:A cholesteryl ester transfer complex in human plasma. Proc. Natl. Acad. Sci. U.S.A., 77:3327 (1980). 18) Albers, J. J., Tollefson, J. H., Chen, C-H. and Steinmetz, A.:Isolation and characterization of human plasma lipid transfer proteins. Arteriosclerosis, 4:49 (1984). 19) Barter, P. J., Gooden, J. J. M., Rajaram, O. V.: Species differences in the activity of a serum triglyceride transfering factor. Atherosclerosis, 33:165 (1979). 20) Oschry, Y. and Eisenberg, S.:Rat plasma lipoproteins:re-evaluation of a lipoprotein system in an animal devoid of cholesteryl ester transfer activity. J. Lipid Res., 23:1099 (1982). 21) Barter, P. J., Ha, Y. C. and Calvert, G. D.:Studies of esterified cholesterol in sub-fractions of plasma high density lipoproteins. Atherosclerosis, 38:165 (1981).
9 22) Fielding, C. J., Reaven, G. M., Liu, G. and Fielding, P. E.:Increased free cholesterol in plasma low and very low density lipoproteins in non-insulin-dependent diabetes mellitus:its role in the inhibition of cholesteryl ester transfer. Proc. Natl. Acad. Sci. U.S.A., 81:2512 (1984). 23) Breckenridge, W. C., Little, J. A., Alaupovic, P., Wang, C. S., Kuksis, A., Kakis, G., Lindgren, F. and Gardiner, G.:Lipoprotein abnormalities associated with a familial deficiency of hepatic lipase. Atherosclerosis, 45:161 (1982). 24) Gordon, V., Innerarity, T. L. and Mahley, R. W.: Formation of cholesterol- and apoprotein E- enriched high density lipoproteins in vitro. J. Biol. Chem., 258:6202 (1983). 25) Miettinen, T. A.:Mode of action of a new hypocholesterolemic drug (DH-581) in familial hypercholesterolemia. Atherosclerosis, 15:163 (1972). 26) Nestel, P. J. and Billington, T.:Effects of probucol on low density lipoprotein removal and high density lipoprotein synthesis. Atherosclerosis, 38: 203 (1981). 27) Kesaniemi, Y. A. and Grundy, S. M.:Influence of probucol on cholesterol and lipoprotein metabolism in man. J. Lipid Res., 25:780 (1984).
10 Clinical and Biochemical Studies on Hyper-HDL2-cholesterolemia Associated with Juvenile Corneal Opacity Shlzuya YAMASHITA*, Toshiharu KAWAMOTO*, Tadashl NAKAMURA*, Shuichi NOZAKI*, Tohru FUNAHASHI*, Sheng CHIAO*, Shigenor i FUJIOKA*, Tadahisa NAKAJIMA*, Hiroshi KAMIDO*, Kaoru KAMEDA*, Kazuhiko HIROBE*, Yuji MATSUZAWA*, Mltsuyo OKAZAKI**, Ichiro HARA***, Shinji YOKOYAMA****, Akira YAMAMOTO**** and Sei ichiro TARUI* *Second Department of Internal Medicine, Osaka University Medical School Fukushima-ku, Osaka 553, Japan **Laboratory of Chemistry, Department of General Education, Tokyo Medical and Dental University, Kohnodai, Ichikawa, Chiba 272, Japan ***Toyo Soda Co., Tokyo 107, Japan ****Department of Etiology and Pathophysiology, National Cardiovascular Center Research Institute, Suita, Osaka 565, Japan We recently discovered two homozygous cases of hyper-hdl2-cholesterolemia associated with premature corneal opacities. Patient 1 is a 55- year-old man, whose serum HDL-cholesterol levels were much elevated to -140mg/dl, while serum total cholesterol was only slightly increased and serum triglyceride moderately reduced. Serum levels of apolipoprotein (apo) A-I and E were also elevated. Patient 2 is a 58-year-old man whose serum HDL-cholesterol was also much increased to mg/dl. The lipid and lipoprotein patterns of these patients were closely similar by ultracentrifugal analysis and the increase of HDLcholesterol was attributed solely to the enrichment of cholesteryl ester (CE) in HDL2 fraction (1.063< d<1.125g/ml). The trait was considered to be inherited in the family of Patient 2. In postheparin plasma, LPL activities were increased in these patients, whereas H-TGL showed reduced activities. In order to elucidate the mechanism of accumulation of CE in HDL2, we examined CE net transfer rate according to the modified method of Fielding et al. In control subjects there was a net mass transfer of CE from HDL to VLDL and LDL after a 1 hr's incubation with DTNB at 37 Ž, while in both cases there was a net transfer of CE in the opposite direction leading to the accumulation of CE in HDL. We also examined the effect of probucol on the lipoprotein and apoprotein levels of these patients. Probucol reduced serum levels of HDL-cholesterol, apo A-I and apo E parallel to the reduction of serum total cholesterol. The decrease of HDLcholesterol was largely due to the reduction of cholesterol in HDL2 fraction. Examination of patients' sera by HPLC disclosed that large HDL2 particles became smaller to almost normal sizes after the treatment with probucol and that the change was proved to be reversible after stopping administration of the drug. We also examined CE net transfer rate after treatment with probucol, and found that the reversion of CE net transfer was normalized in both cases after the therapy. We therefore conclude that abnormalities in CE net transfer might be involved in the pathogenesis of hyper-hdl2-cholesterolemia. Key wards:hyper-hdl 2-cholesterolemia, Familial, Corneal opacity, Cholesteryl ester transfer rate, Probucol.
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