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J K Choi

Publications and source records attributed to J K Choi.

85 records · Page 5Linked to original sources

Structure of the horseradish peroxidase isozyme C genes.

We have isolated, cloned and characterized three cDNAs and two genomic DNAs corresponding to the mRNAs and genes for the horseradish (Armoracia rusticana) peroxidase isoenzyme C (HPR C). The amino acid sequence of HRP C1, deduced from the nucleotide sequence of one of the cDNA clone, pSK1, contained the same primary sequence as that of the purified enzyme established by Welinder [FEBS Lett. 72, 19-23 (1976)] with additional sequences at the N and C terminal. All three inserts in the cDNA clones, pSK1, pSK2 and pSK3, coded the same size of peptide (308 amino acid residues) if these are processed in the same way, and the amino acid sequence were homologous to each other by 91-94%. Functional amino acids, including His40, His170, Tyr185 and Arg183 and S-S-bond-forming Cys, were conserved in the three isozymes, but a few N-glycosylation sites were not the same. Two HRP C isoenzyme genomic genes, prxC1 and prxC2, were tandem on the chromosomal DNA and each gene consisted of four exons and three introns. The positions in the exons interrupted by introns were the same in two genes. We observed a putative promoter sequence 5' upstream and a poly(A) signal 3' downstream in both genes. The gene product of prxC1 might be processed with a signal sequence of 30 amino acid residues at the N terminus and a peptide consisting of 15 amino acid residues at the C terminus.

Amino Acid Sequence↗

Changes in renal function in cadmium-intoxicated rats.

Changes in renal function, Na+-K+-ATPase activity and PAH transport system in kidney cortex were studied in rats treated with cadmium. Subcutaneous injections of CdCl2 (2 mg Cd/kg.day) for 16 days induced a marked polyuria and a hyposthenuria. These changes were accompanied by increase in urinary protein, glucose, urea, calcium, phosphate, chloride and potassium excretions. The change in urine flow was proportional to the change in total osmotic solute excretion. Creatinine excretion and TcH2O remained unchanged. Na+ excretion was not increased, but the Na+-K+-ATPase of renal cortex was significantly inhibited. PAH uptake by renal cortical slices was markedly attenuated in Cd-treated rats. The Vmax for active PAH influx was drastically reduced, but the Km was not changed. The passive influx and efflux of PAH across the basolateral membrane and the renal tissue oxygen consumption were not apparently altered in Cd-treated animals. These results indicate that 1) the nature of Cd-induced polyuria and hyposthenuria is an osmotic diuresis induced by proximal tubular rejection of various substances, and 2) the mechanism of impaired renal PAH excretion in Cd-treated animals is a loss of organic anion carriers in proximal tubular basolateral membranes.

Animals↗

Effect of wearing gloves on the thermal balance of Korean women wet-suit divers in cold water.

Effect of wearing neoprene gloves on the thermal exchanges of wet-suited divers was studied in 8 Korean diving women. Subjects, clad with 5-6-mm-thick neoprene wet suits (jacket, pants, and boots) either with or without wearing 3-mm-thick neoprene gloves, were immersed for 3 h in water of critical temperature (17.3 degrees +/- 0.8 degree C) while the rectal and skin (chest, leg, arm, and hand) temperatures and oxygen consumption were measured. Overall thermal insulation of the subject plus suit was calculated from the rectal-to-water temperature difference divided by the estimated rate of skin heat loss. The skin heat loss was assumed to equal metabolic heat production minus respiratory heat loss, corrected for changes in heat storage when mean body temperature changed. All measurements were carried out in a resting condition. During the 3rd h of immersion, the rectal temperature was lower with gloves (delta Tre = 0.30 degree +/- 0.04 degree C; P less than 0.05) whereas metabolic heat production was not significantly different. Consequently, the total thermal insulation was nearly 16% lower with gloves than without gloves. In both the hands and forearms, the regional heat flux determined directly using a heat flux transducer was higher and the thermal insulation index was lower with gloves than without gloves. These results indicate that in wet-suited subjects resting in cold (17 degrees C) water gloves do not provide additional protection against heat loss, but rather decrease the efficiency of thermoregulatory mechanisms. We suggest that sensory input from cold receptors in the distal extremities is particularly important in thermoregulation during immersion in cold water.

Adult↗

Changes in thermal insulation during underwater exercise in Korean female wet-suit divers.

The present work was undertaken to examine the effect of wet suits on the pattern of heat exchange during immersion in cold water. Four Korean women divers wearing wet suits were immersed to the neck in water of critical temperature (Tcw) while resting for 3 h or exercising (2-3 met on a bicycle ergometer) for 2 h. During immersion both rectal (Tre) and skin temperatures and O2 consumption (VO2) were measured, from which heat production (M = 4.83 VO2), skin heat loss (Hsk = 0.92 M +/- heat store change based on delta Tre), and thermal insulation were calculated. The average Tcw of the subjects with wet suits was 16.5 +/- 1.2 degrees C (SE), which was 12.3 degrees C lower than that of the same subjects with swim suits (28.8 +/- 0.4 degrees C). During the 3rd h of immersion, Tre and mean skin temperatures (Tsk) averaged 37.3 +/- 0.1 and 28.0 +/- 0.5 degrees C, and skin heat loss per unit surface area 42.3 +/- 2.66 kcal X m-2 X h. The calculated body insulation [Ibody = Tre - Tsk/Hsk] and the total shell insulation [Itotal = (Tre - TW)/Hsk] were 0.23 +/- 0.02 and 0.5 +/- 0.04 degrees C X kcal-1 X m2 X h, respectively. During immersion exercise, both Itotal and Ibody declined exponentially as the exercise intensity increased. Surprisingly, the insulation due to wet suit (Isuit = Itotal - Ibody) also decreased with exercise intensity, from 0.28 degrees C X kcal-1 X m2 X h at rest to 0.12 degrees C X kcal-1 X m2 X h at exercise levels of 2-3 met.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Identification of a chymotrypsin-like proteinase in human mast cells.

An antiserum was produced against a chymotryptic proteinase purified from human skin. The antiserum did not cross-react with human leukocyte cathepsin G and elastase, rat mast cell proteinase I, and human skin tryptase. Indirect immunofluorescent staining of frozen skin sections to localize the proteinase showed cytoplasmic staining of cells scattered about the papillary dermis and around blood vessels and appendages. Restaining these sections with toluidine blue revealed that the fluorescently stained cells contained metachromatically staining granules, the major distinguishing feature of mast cells. A similar correlation was found in lung tissue. Ultrastructural studies employing the ferritin bridge technique to immunologically identify the proteinase additionally localized the proteinase to mast cell granules. Biochemical and immunochemical characterization of chymotryptic activity solubilized from isolated human lung mast cells identified a chymotryptic proteinase that may be identical to the skin chymotryptic proteinase. These studies establish that human skin mast cells contain a chymotrypsin-like proteinase that is a granule constituent and provide evidence that indicates a comparable proteinase is also present in lung mast cells.

Chymases↗

Thickening of the capillary basement membrane of the quadriceps muscle in Vacor-induced diabetic patients.

The effect of Vacor diabetes mellitus on the quadriceps muscle capillary basement membrane thickness has been examined by the electron microscopic method. With this procedure, it was possible to obtain an objective evaluation of the significance of capillary basement membrane thickening in Vacor diabetes mellitus. The results of such a study on the muscle capillary basement membrane in 10 normal subjects and 9 Vacor-poisoned patients have demonstrated the following: (1) the average basement membrane width of Vacor patients (2801 A +/- SD 614) is over twice that of normal subjects (1300 +/- 185 A), and all 9 cases demonstrated muscle capillary basement membrane thickening; (2) there is a significant association of basement membrane width with age (P less than 0.01), but not with the duration of the disease or fasting blood glucose values in the Vacor group. The results of the present study indicate that diabetes mellitus induced by Vacor poisoning can lead to microangiopathy, and that such basement membrane thickening is an early and consistent finding.

Adolescent↗

Expression of p50 C-terminal Src kinase (Csk) in mouse testis.

C-terminal Src Kinase (Csk) is a cytoplasmic tyrosine kinase that phosphorylates a critical tyrosine residue in each of the Src family kinases to inhibit their activities. To investigate the possible regulation of spermatogenesis by Src-Csk loop, the postnatal changes in the expression of Csk were examined in mouse testes. Semiquantitative RT-PCR analysis revealed that Csk mRNA increased during neonatal development and peaked at 2 weeks of age. Following the decrease during pubertal development, Csk expression re-increased in adult testes. In Western blot, immature testes showed higher expression of Csk protein than the pubertal or adult testes. In immature testis, Csk immunoreactivity was largely found in the Sertoli cell and there was no visible difference in the Csk immunoreactivity among the seminiferous tubules. In adult testis, however, a differential Csk immunoreactivity was found among the seminiferous tubules. Intense signal was found in the adluminal cytoplasm of the Sertoli cells bearing the post-meiotic differentiating germ cells, suggesting that Csk may participate in the remodeling of seminiferous tubule during late phase of spermatogenesis. Csk immunoreactivity was also found in the Leydig cells, suggesting the possible regulation of Leydig cell function. Src-Csk loop may participate in the differentiation of the seminiferous epithelia and Leydig cells in mouse testis.

Aging↗

Chemical degradation of tobacco mosaic virus followed by infectivity assay, reverse transcription-polymerase chain reaction and gel electrophoresis.

In order to determine the detection limit for chemically treated virions by gel electrophoresis, reverse transcription-polymerase chain reaction (RT-PCR) and infectivity assay, tobacco mosaic virus (TMV) exposed to various concentrations of chemicals was studied. When virions were exposed to 0.2 N HCl for 30 mins, partially degraded TMV particles were observed by gel electrophoresis. Under the same exposure, a major RT-PCR amplified DNA product corresponding to the target size of 806 bp, which decreased as a function of time, could be detected for up to 60 mins of exposure. When virions were treated with NaOH (0.02 N or higher normality) for 5 mins, partially degraded virions were detected by gel electrophoresis, exhibiting multiple band patterns. Exposure of the virions to 0.1 N NaOH for 5 mins revealed severely degraded viral RNA, but disappearance of the amplified RT-PCR products was apparent during 30-60 mins of exposure. Therefore, these data showed clearly the difference in the detection limit of gel electrophoresis and that of RT-PCR for the degraded viral RNA. In addition, the infectivity assay showed that the number of local lesions in Nicotiana rustica were significantly reduced by more than 95% when the virus was exposed to 0.2 N HCl for 15 mins or 0.1 N NaOH for 10 mins. From these results we conclude that loss of infectivity was not related to that of PCR product. Other chemical disinfectants such as phenol or formalin were also found to be effective to reduce the virus infectivity, but a corresponding degradation of viral RNA was detected by neither gel electrophoresis nor RT-PCR.

Electrophoresis, Agar Gel↗

Intranuclear post-transcriptional down-regulation responsible for loss of a keratin differentiation marker in tumour progression.

Apparent loss of differentiation markers characterizes advanced malignant neoplasms. Post-transcriptional down-regulation of keratin message to levels undetectable with a partial cDNA probe to rat keratin K5 had been observed in anaplastic cells (T952/F7) derived from benign keratin-producing cells (A5P/B10) (1). The entire fifth introns of both the K5 and K6 genes were generated from rat genomic DNA by PCR to define expression of these closely related proteins. Sequencing of the PCR products revealed 84% homology in the K5 and K6 exon regions included, but absence of any homology in the introns. Active transcription of K5 could be demonstrated in the anaplastic cells with reverse transcription of nuclear RNA (RTn-PCR) by the presence of PCR-generated products confirmed by sequencing as unspliced and spliced transcripts of rat K5. In situ hybridization with ssDNA probes for the spliced message from this region of the K5 gene demonstrated a punctuate distribution in the cytoplasm of the benign cells and absence of any detectable message in the anaplastic derivatives, ssDNA probes for the unspliced transcript containing intron 5 and the same flanking exon sequences as the spliced probe detected transcription of hnRNA in the anaplastic cells as discrete signals confined to the nuclear compartment. These results show that failure to express mRNA for a differentiation marker in the cytoplasm of anaplastic cells can be due to a mechanism operating in the nuclear compartment after gene transcription and indicate that the mechanism functions shortly after splicing of the transcript.

Animals↗