According to the authors' clinical analysis, about half of the patients who suffer from arteriosclerotic obstruction (ASO) in the lower extremity(-ies) with clinical manifestation are dyslipidemic (total cholesterol > or = 220 mg/dL or LDL cholesterol > or = 140 mg/dL). As suggested by clinical success in regression of ASO in the coronary arteries as a result of aggressive removal of LDL, LDL adsorption utilizing an extracorporeal circulation technique with a dextran sulfate/cellulose adsorbent column was applied in 33 patients (22 men and 11 women) with ASO. Clinical results obtained after a series of 10 LDL adsorption procedures as a standard showed encouraging success. Improvement in subjective symptoms was achieved as follows: 88.5% for cold lower extremity, 87.1% for intermittent claudication, 53.8% for leg/toe pain at rest, and 60% for disappearance/size diminution of ulcer/necrosis. Improvements in objective examination findings supported subjective ones: 85.7% by plethysmography, 81% by thermography and 70% by ankle pressure index. No serious complications or untoward effects were observed during or after the adsorption procedures. In conclusion, LDL adsorption appears to be a useful and safe tool in treatment of ASO patients with dyslipidemia.

DL-threo-and DL-erythro-Cn-dihydrosphingosines (n=12, 14, and 16) were separated from the corresponding 2-amino-1, 3-alkanediols by repeated recrystallization of the N, O, O-triacetyl and N-acetyl derivatives. Ceramides having DL-threo-and DL-erythro-Cn-dihydrosphingosines were prepared by reactions of the bases with N-acyloxysuccinimides in tetrahydrofuran. CLC-MS analysis of O, O-di-TMS derivatives of the ceramides was carried out to find differences between threo and erythro isomers in mass spectra. One characteristic difference was observed in the relative intensity (RI) at m/e=M-103; i.e., the relative intensities of threo compounds (RI 40•`80%) were stronger than those of erythro isomers (RI 6•`14%). The ratios, RI(M-103)/RI(M-15), were 3.7•`6.8 and 0.8•`1.3 for threo and erythro compounds, respectively. These observations demonstrate the possibility of determining erythro and threo isomers by mass spectrometry.

Yeast ENO1 promoter was prepared by a chemical synthetic method. Two variant promoters containing a pyrimidine-rich region (CT block), located between the TATA box and the transcription start site, either 32 or 34 base pairs (bp) longer than the native ENO1 promoter were isolated during the chemical synthesis. Gene expression of variant promoters was compared with that of the native promoter by measuring the amount of mRNA and the activity of beta-galactosidase by constructing ENO1-lacZ gene fusions. No significant differences were observed between the native and variant promoters in transcription levels. The start site of transcription was mapped on CAAG, a consensus sequence of transcription start site of yeast glycolytic genes. The results suggest a longer CT block in ENO1 promoter may not affect the expression of the yeast ENO1 gene. In addition, the level of ENO1 gene expression was found to be higher in stationary phase cells than in log phase cells.

Several plasmids carrying different length of the 5' flanking region of yeast (Saccharomyces cerevisiae) enolase 1 gene (ENO1) which is fused in frame to the Escherichia coli lacZ gene were constructed by recombination in vitro. Promoter activity of ENO1 was assayed by measuring beta-galactosidase activity of the fused gene product. Comparison of the promoter activity of these plasmids suggests that the sequences required for a strong promoter activity lie within the DNA segment -724 to -353 base pairs (bp) upstream from the start of ENO1 coding sequence. The nucleotide sequence of this region was determined.