Biomedical subjects
Publications and source records attributed to K Fujiyama.
Human growth hormone gene with 584 base pairs has been synthesized by joining with DNA ligase of chemically synthesized deoxyoligonucleotides with the chain length of 15-25. The phosphotriester polymer support synthesis was employed to obtain 80 oligonucleotides.
An E. coli tRNAfMet fragment [C-A-U-A-A-C-C-C-G-A-A-G-G-U-C-G-U-C-G-G (bases 35-f54)] containing the anticodon triplet has been synthesized by the phosphotriester method involving protected oligonucleotide blocks. Di- or tri-nucleotide blocks were prepared by condensation of 2'-O-(o-nitrobenzyl) nucleotide derivatives and used for the synthesis of pentanucleotide blocks. The 5'-hydroxy, heterocyclic amino and internucleotide linkage were protected with monomethoxytrityl, acyl and p-chlorophenyl groups, respectively. The 3'-phosphates of the pentanucleotides, except for the GUCGG block where 2'-O-benzoyl 3'-O-(o-nitrobenzyl) N-isobutyrylguanosine was used, were protected with p-chlorophenyl and anilido groups. The anilido groups were removed by treatment with isoamyl nitrite and the 3'-phosphodiesters of resulting pentamers were activated with mesitylenesulfonyl nitrotriazolide to give protected decanucleotides in yields of 61-89%. The two decanucleotides were condensed similarly to yield the protected eicosanucleotide in a yield of 59%. The product was deblocked and purified by ion-exchange chromatography on DEAE-Sephadex A-25 and characterized by enzymatic hydrolysis after labelling the 5'-end by phosphorylation using polynucleotide kinase and [gamma-32P]ATP.
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Oligonucleotides corresponding to the E. coli tRNAfMet fragments and their analogs have been synthesized by the phosphodiester or phosphotriester method. The nascent fragments were joined by using RNA ligase to yield a total tRNA molecule. The synthetic molecule was tested its ability to accept methionine in the presence of purified methionyl-tRNA synthetase from E. coli. The charged tRNA and the hydrolyzed methionine were isolated by gel filtration. The discharged tRNA was characterized by nearest neighbor analysis and 5'-end analysis.
Oligonucleotides corresponding to the total tRNAfMET FROM E. coli have been synthesized. These fragments were joined by using RNA ligase to yield quarter molecules. The 5'-quarter molecule showed 84% amino acid acceptor activity when it was combined with the natural three quarter molecule.
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To elucidate the changes of serum apolipoproteins as possible atherogenic factors in non-insulin-dependent diabetes mellitus (NIDDM), comparison was made between 47 NIDDM patients with clinical macroangiopathy (coronary heart disease and/or cerebral infarction) and 47 NIDDM patients without clinical macroangiopathy individually matched for age, sex, body mass index, glycemic control, mode of therapy, and blood pressure. Serum total cholesterol (223 +/- 63 mg/dl, mean +/- SD) and triglyceride (160 +/- 72 mg/dl) levels in NIDDM with clinical macroangiopathy were significantly higher than those (198 +/- 44 and 126 +/- 78 mg/dl) in control NIDDM. Apolipoprotein AII (apo AII) (28.8 +/- 6.9 mg/dl) in NIDDM with clinical macroangiopathy was significantly lower than that (31.4 +/- 5.5 mg/dl) in control NIDDM, and apo B (147 +/- 48 mg/dl) and apo CIII (13.1 +/- 5.9 mg/dl) in NIDDM with clinical macroangiopathy was significantly higher than that (113 +/- 35 and 10.8 +/- 4.7 mg/dl) in control NIDDM, respectively. There were no significant differences in apo AI, CII, and E levels. The ratio of HDL cholesterol to apo AI (0.36 +/- 0.08) and total cholesterol to apo B (1.6 +/- 0.3) in NIDDM with clinical macroangiopathy was significantly lower than that (0.40 +/- 0.09 and 1.8 +/- 0.4) in control NIDDM. These results suggest that abnormality of lipid metabolism is more significant in NIDDM with clinical macroangiopathy than in NIDDM without clinical macroangiopathy, and is more pronounced in women than in men.