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Biomedical subjects

S C Kim

Publications and source records attributed to S C Kim.

At least 289 records · Page 16Linked to original sources
PubMed1986

Isotopic probes of the argininosuccinate lyase reaction.

The mechanism of the argininosuccinate lyase reaction has been probed by the measurement of the effects of isotopic substitution at the reaction centers. A primary deuterium isotope effect of 1.0 on both V and V/K is obtained with (2S,3R)-argininosuccinate-3-d, while a primary 15N isotope effect on V/K of 0.9964 +/- 0.0003 is observed. The 15N isotope effect on the equilibrium constant is 1.018 +/- 0.001. The proton that is abstracted from C-3 of argininosuccinate is unable to exchange with the solvent from the enzyme-intermediate complex but is rapidly exchanged with solvent from the enzyme-fumarate-arginine complex. A deuterium solvent isotope effect of 2.0 is observed on the Vmax of the forward reaction. These and other data have been interpreted to suggest that argininosuccinate lyase catalyzes the cleavage of argininosuccinate via a carbanion intermediate. The proton abstraction step is not rate limiting, but the inverse 15N primary isotope effect and the solvent deuterium isotope effect suggest that protonation of the guanidino group and carbon-nitrogen bond cleavage of argininosuccinate are kinetically significant.

Animals↗
PubMed1986

The determination of enzyme-substrate dissociation rates by dynamic isotope exchange enhancement experiments.

A new method for the determination of dissociation rates of enzyme-substrate complexes has been developed. The rate of exchange of a labeled product back into the substrate is measured during catalysis of the forward reaction when the forward reaction is kept far from equilibrium by the enzymatic removal of the nonexchanging product. The ratio of the exchange rate and the net rate for product formation is then determined at various concentrations of the exchanging product. A plot of this ratio is a diagnostic indication of the kinetic mechanism and the relative rates of product dissociation from the binary and ternary enzyme complexes. This technique has been applied to the reaction catalyzed by bovine liver argininosuccinate lyase. The ratio for the rate of exchange of fumarate into argininosuccinate and the net rate for product formation was found to increase with the concentration of fumarate but to reach a limit of 3.3. The ratio of rates was half-maximal at 36 mM fumarate. The data have been interpreted to indicate the argininosuccinate lyase has a random kinetic mechanism. The calculated lower limit for the rate of release of arginine from the enzyme-fumarate-arginine complex is 0.35 times as fast as the Vmax in the reverse direction. The rate of release of arginine from the enzyme-arginine binary complex is 210 times faster than Vmax in the reverse direction.

Animals↗
PubMed1986

A novel multistep method for generating precise unidirectional deletions using BspMI, a class-IIS restriction enzyme.

A novel approach is described that permits the introduction of unidirectional deletions into a cloned DNA fragment, in a precisely controlled manner. The method is based on the use of a special vector and a class-IIS restriction endonuclease, BspMI, which produces staggered cuts 4 and 8 nucleotides (nt) to the 3' from its recognition site 5'-ACCTGC-3'. The DNA fragment is inserted into the pUC19-based plasmid, which contains a unique BspMI recognition site, and the appropriate number of cleavage-and-deletion cycles is performed, each cycle removing 4 bp. Since the recognition site is not affected by the BspMI cleavage, no recloning of the DNA fragment is necessary. Each cycle consists of BspMI cleavage, removal of the 4-nt single-stranded cohesive ends with mung bean nuclease (MB), and blunt-end ligation to recircularize the plasmid. The shortened plasmid is reintroduced into the host, after one or after several such 4-bp deletion cycles. When DNA is inserted into the multiple cloning site in the lacZ alpha gene, the progress of 4-bp removal can be followed by determining the Lac phenotype, since removal of multiples of 3 bp retains the reading frame while other kinds of deletions distort (or restore) the reading frame. Loss of pre-existing restriction sites or creation of new ones also permits monitoring the progress of the deletion process.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromosome Deletion↗
PubMed1983

Symptomatic neurosyphilis.

We observed 24 patients with symptomatic neurosyphilis. Of these 24 patients, nine had clinical manifestations of general paresis, seven tabetic neurosyphilis, five meningovascular neurosyphilis, one taboparetic, one tabomeningovascular, and one spinal neurosyphilis. Confusion, disorientation, lancinating pain, impaired memory, and bizarre behavior were the most prevalent symptoms. Sensitivity of the CSF FTA-ABS test was higher (96%) than that of the CSF VDRL test (79%). CSF cell count and total protein determination may be inadequate for assessing activity of neurosyphilis.

Adult↗
PubMed1980

Correlation of cell cycle parameters with radiation sensitivity in a series of murine L5178Y cells.

The X-ray dose--response parameters (Do, n and Dq) of 10 strains of murine L5178Y cells spanning a wide range of radiation sensitivities were characterized. The proportions of cells residing in G1, S and G2 + M phases of the cell cycle in exponentially growing cell populations were estimated using DNA microfluorometric techniques. The radiosensitive strains contained a lower proportion of cells in G1 phase and a somewhat larger proporation of cells in S phase than did the radioresistant strains, reflecting a direct correlation between radioresistance and the duration of G1 phase (and possibly an inverse correlation with the duration of the S phase) in these closely related cell strains. These results are discussed in terms of the putative role of G1 in governing radiosensitivity of cells and tumours.

Animals↗
PubMed1980

Local anaesthetics as hypoxic radiosensitizers, oxic radioprotectors and potentiators of hyperthermic killing in mammalian cells.

Local anaesthetics with well-known pharmacology are found to radiosensitize hypoxic and radioprotect oxic murine L5178Y cells. Under the specified experimental conditions, radiation-modifying effects exerted by local anaesthetics are dose-dependent. It appears that in order that the drugs should act as oxic radioprotectors or as hypoxic radio-sensitizers, the agent must be present during the period of irradiation. Local anaesthetics are also found to potentiate the hyperthermic killing of cells. The possibility of applying compounds with such dual radiation-modifying properties plus hyperthermia-potentiating capacity to enhance the therapeutic gains in tumour therapy is briefly discussed.

Anesthetics, Local↗
PubMed1979

Selection and characterization of a varient of murine L5178Y lymphoma resistant to local anesthetics.

A varient of murine L5178Y lymphoma resistant to procaine hydrochloride (PH) was selected by exposing the cells to gradual increments of PH in the growth medium until the cell grew exponentially in the presence of 1.5 mM PH. Using cinephotomicrography, it was observed that the majority of cells that initially succumbed to PH failed to undergo successful mitosis. With respect ot chromosomal, cell size distribution and flow microfluorometric analyses, the PH-resistant cells are very similar to a spontaneous tetraploid cell line (R1T) previously cloned. The isolated cells, designated R1/P, were also found to be cross-resistant to analogues of PH, namely, lidocaine, tetracaine and dibucaine. The naturally-occurring tetraploid cell line (R1T) was also found to be more resistant to local anesthetics, although not to the same extent as R1/P cells. Since the enzyme that hydrolyzes procaine appears to be absent in all these lymphoid cell lines, the difference in resistance does not appear to depend on differences in the ability of these cells to remove the agent. It is suggested that an alteration in the structure and/or function of the plasma membrane in R1/P cells have rendered them either less sensitive to the membrane-perturbing effects of the local anesthetics or less permeable to local anesthetics molecules. The ability of local anesthetics to affect membranes and cytoskeleton structures may play a role in the genesis and/or selection of these cell variants.

Animals↗
PubMed1979

Effect of low dose rate radiation on cell growth kinetics.

Experimental determinations were made of cell number as a function of time for two strains of L5178Y mammalian cells maintained continuously in various environments of radiation. One strain possessed a shoulder in its dose response curve whereas the other did not. Neither strain showed any significant difference in growth rate for interdivision doses on the order of the median lethal dose or less delivered continuously at a low dose rate or pulsed every 4 h at a high instantaneous dose rate. It was also shown that large numbers of dead cells have little effect on growth rate and that these dead cells last as discrete entities for many days. A simple theory of growth rate in the presence of radiation is presented, and the agreement with the observations implies that there is no effect of any sublethal low dose rate radiation received in one generation on the growth rate or radiation sensitivity of the succeeding generation. Further analysis of the data also showed that for the no-shoulder cells at 37 degrees C, tritiated water had a relative biological effect close to unity for cell sterilization.

Animals↗