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Biomedical subjects

T Doi

Publications and source records attributed to T Doi.

At least 505 records · Page 28Linked to original sources

Total synthesis of tRNAfMet.

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.

Base Sequence↗

A new method for 3'-labelling of polyribonucleotides by phosphorylation with RNA ligase and its application to the 3'-modification for joining reactions.

P1-Adenosine 5'-P2-o-nitrobenzyl pyrophosphate (nbzlppA) has been synthesized as a substrate for T4 RNA ligase catalyzed 3'-phosphorylation. Incubation of oligoribonucleotides and nbzlppA with RNA ligase yielded oligoribonucleotides having a 3'-o-(o-nitrobenzyl) phosphate. Photochemical removal of the o-nitrobenzyl group provided the free 3'-phosphate. Using [P2-32P] nbzlppA, 3'-termini of oligoribonucleotides could be labelled with 32P. This reaction was applied to modify the 3'-end of donor molecules in joining reaction with RNA ligase. A trinucleotide U-A-G was converted to U-A-Gpnbzl and phosphorylated with polynucleotide kinase. pU-A-Gpnbzl was then joined to an acceptor trinucleotide A-U-G to yield A-U-G-U-A-Gp.

Adenosine Diphosphate↗

Synthesis of the nascent strand of tRNAfMet from E coli.

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.

Base Sequence↗

The role of OEP-antibodies in Pseudomonas aeruginosa infection.

When mice were challenged intraperitoneally with 2.2 X 10(4) Pseudomonas aeruginosa in 0.5 ml of 2.7% mucin solution, the bacteria markedly increased in the peritoneal cavity, rapidly entered the blood stream, and the mice died, probably of bacteremia, within a short time. However, when mice were injected subcutaneously 4 hours prior to the challenge with 0.2 ml of 0.75% IgG (OEP-HA = 12.8) or IgM (OEP-HA = 64) separated from human plasma or 3% S-IgA EP-HA = 19.2) from fresh human milk, the bacteria were inhibited from propagating and were eliminated from the cavity so that they could not enter the blood stream. As a result, the mice tolerated the challenge and survived. When 3% IgA (OEP-HA less than 0.8) from human plasma was given, a complete cure was not effected in a few mice despite a general tendency of healing. Determination of the number of viable intra- and extracellular bacteria in the peritoneal cavity revealed that the infecting bacteria were rapidly phagocytized and killed by the peritoneal phagocytes in the presence of OEP-antibodies. These findings indicate that the effect of bacterial clearance from the cavity results from the cooperative action of OEP-antibodies and peritoneal phagocytes. Marked enhancement of phagocytosis by the antibodies was observed in an in vitro study in the presence of a heat-labile, complement-like substance(s). This finding is interpreted as evidence of the cooperative activity.

Animals↗

Homovanillic acid in human cerebrospinal fluid.--Its concentration gradient and reduced levels in patients with epilepsy.

The homovanillic acid (HVA) concentrations in the lumbar cerebrospinal fluid (CSF) were determined in 38 epileptic and 39 control patients. The mean concentration of HVA was 23.9 ng/ml +/- 2.8 SEM for the epileptic group and 30.2 ng/ml +/- 2.1 SEM for the control group, respectively. Thus, HVA was significandly reduced in the patients with epilepsy compared with the controls. The mean HVA in the female patients was higher than in the male patients in both groups but this failed to reach statistical significance. There was no apparent relationship between the degree of reduced HVA concentration and other clinical indexes of the epilepsy (age, type and frequency of seizures, and anticonvulsant medication). For the determination of concentration gradient of HVA three fractions of the spinal CSF were obtained from 11 patients. A pronounced gradient of HVA concentration was found with a ratio of 1 : 1.46 : 1.97 for the first, second and third fractions. This suggests that a standardized conditions for collecting CSF should be employed to study HVA levels in humans.

Adult↗