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Y Umezawa

Publications and source records attributed to Y Umezawa.

8 recordsLinked to original sources

Efficient cDNA cloning method using solid phase DNA probe.

We are now developing a novel and efficient method using solid phase DNA probe to isolate a particular recombinant cDNA from single stranded cDNA library. Target clone coding metapyrocatechase (MPC) and cDNA library constructed from mRNA of U-937 (human lymphoma cell line) were converted to single stranded form by superinfection of helper phage (M13KO7). Probe DNA (25 mer) composed of a portion of the target cDNA was synthesized, attached to an HPLC gel and used as a solid phase DNA probe. Hybridization between probe DNA and target clone was performed in an Eppendorf tube within a few hours. Competent cell (JM109) was transformed with about one-twentieth of hybridized and eluted fraction by Hanahan's method. From the mixture of 1 ng of MPC vector and 5 micrograms of cDNA library, we obtained 50 colonies containing MPC gene out of 63 transformed colonies.

Catechol 2,3-Dioxygenase

Polygraphic study on the clinical seizures induced during nocturnal sleep.

The subjects of this study consisted of 17 epileptic patients with clinical seizures during all-night sleep recording (the seizure group) and another 17 epileptic patients without clinical seizures (the control group). The results obtained were as follows: 1) The 50-90% out of total clinical seizures were induced in non-REM sleep, while a few clinical seizures were induced in REM sleep. The number of clinical seizures in sleep stage of higher activity level increased as the incidence of clinical seizure during all-night grew. 2) In the seizure group REM sleep could not be detectable during all-night in five cases out of 17 cases, while we could not find cases lacking REM sleep in the control group. This difference between two subject groups was statistically significant (P less than 0.025). 3) As to focal spike group, the spike discharge incidence of total sleeping time and of each sleep stage was higher respectively in the seizure group than that in the control group, and particularly in St.1, St.2 and REM sleep the figure of the discharge incidence was found significantly higher in the seizure group than that in the control group (P less than 0.05). We discussed on above-mentioned results, and we emphasized particularly that REM-sleep suppresses clinical seizures, although non-REM sleep induces clinical seizures.

Adolescent

Aminoglycoside 3'-phosphotransferase III, a new phosphotransferase. Resistance mechanism.

The aminoglycoside phosphotransferase of Pseudomonas aeruginosa 21-75 was purified by affinity chromatography using dibekacin-Sephadex 4B or lividomycin A-Sepharose 4B followed by DEAE Sephadex A-50 chromatography. It had activities of both the known aminoglycoside 3'-phosphotransferases I and II, and transferred phosphate from ATP to the 3'-hydroxyl group of kanamycin A, ribostamycin and butirosin A and 5-hydroxyl group of lividomycin A. This enzyme was designated aminoglycoside 3'-phosphotransferase III. It showed strong substrate inhibition by kanamycin A and ribostamycin when their concentration exceeded 6 muM. Purification and characterization of this enzyme are reported.

Aminoglycosides