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Biomedical subjects

K C Kim

Publications and source records attributed to K C Kim.

At least 109 records · Page 6Linked to original sources
PubMed1989

Use of primary cell culture to study regulation of airway surface epithelial mucus secretion.

Airway mucus glycoproteins are produced by two different tissues, ie, surface epithelium and submucosa glands. Due to the coexistence of these sources in the same organ, it has been difficult to study the regulation of mucin synthesis and secretion from each source. We have recently developed and characterized a primary culture from tracheal surface epithelium which is highly enriched in secretory cells and produces authentic mucins. Mucins produced by the cultures are extremely hydrophobic. A pool of mucins is localized on the cell surface of secretory cells and these cell surface mucins are released by neutrophil elastase. The simplicity and the relatively rich secretory cell population of the present culture system will serve as a useful model in studying the cell biology of mucin as well as pharmacology of airway surface epithelial mucus secretion.

Ammonia↗
PubMed1988

Characterization of extended primary and secondary cultures of hamster tracheal epithelial cells.

Studies on the regulation of differentiation in airway epithelial cells have been hampered by the lack of cell culture systems that differentiate in vitro. One such system that does exhibit differentiation is hamster tracheal epithelial cells (HTE). A major problem with this system, however, is that at the time cells differentiate, they lyze the collagen gel upon which they grow, resulting in termination of the culture. Here we report that by growing the HTE cells at 32 degrees instead of 37 degrees C we can totally prevent lysis of the collagen gel. Cells grown at this lower temperature maintain their differentiated phenotype as evidenced by abundant mucus granules and the secretion of authentic mucus glycoproteins into the culture media. We have also developed a method for subculturing the primary cells which allows growth and differentiation in secondary culture. The HTE cells were capable of being passaged at least three times and did not become transformed as judged by their inability to grow in soft agar and to produce tumors in syngeneic animals. This improved HTE cell culture system will allow detailed studies on the mechanisms which regulate growth, differentiation, and mucus secretion in surface airway epithelial cells.

Animals↗
PubMed1988

Membrane differentiation markers of airway epithelial secretory cells.

We describe here a system for culturing epithelial cells isolated from hamster trachea, which results in a highly enriched population of mucus-secreting cells. The culture system has enabled us to study the process of secretory cell differentiation in vitro. We found that epithelial secretory cells, in vivo and after 5 days in vitro, selectively bind the lectin Helix pomatia agglutinin (HPA) to apical and, to a lesser extent, basolateral surfaces as well as to mucin granules and intracellular secretory organelles. SDS-PAGE gels of detergent extracts of secretory cells cultured for 5 days reveal three HPA-binding glycoproteins with MW of 120 KD, 220 KD, and greater than 400 KD. The high-MW glycoprotein appears identical to mucin, since it is found in secretions from intact trachea and in spent media from 5-day cultures. It does not appear in spent media from 3-day cultures when cells contain few mucous granules and secrete little mucin. The 220 KD HPA-binding glycoprotein is also present in 5-day but not in 3-day cultures. In contrast, the 120 KD glycoprotein is present at both times. HPA-gp120 is a hydrophobic integral membrane protein, whereas HPA-gp220 and mucin are hydrophilic and are membrane associated. These studies define three membrane glycoproteins, one of which is specific for the tracheal epithelial secretory cell regardless of its mucous content, whereas the other two glycoproteins correlate with mucin secretion. They also demonstrate that, in the fully differentiated state, mucin is bound in a non-covalent fashion to the apical plasma membrane of the tracheal epithelial secretory cell.

Animals↗
PubMed1987

Human neutrophil elastase releases cell surface mucins from primary cultures of hamster tracheal epithelial cells.

Primary hamster tracheal epithelial cells growing on a collagen gel matrix produce high molecular weight mucins indistinguishable from mucins produced in vivo. Using a modified version of these confluent cultures, we have demonstrated here that (i) release of mucins can be stimulated by human neutrophil elastase (HNE; EC 3.4.21.37); (ii) HNE can degrade mucins, and both mucin release and degradation by HNE require an active catalytic site; and (iii) there are at least two pools of mucins in these cells: one is a rapidly turning-over spontaneously releasable constitutive pool, the other is a slowly turning-over HNE-releasable pool. We provide evidence that the HNE-releasable mucins are membrane bound and associated with the secretory cell apical surface.

Animals↗
PubMed1986

Isolation, characterization, and localization of the spanning protein from skeletal muscle triads.

A monoclonal antibody has been developed against the putative junctional protein or spanning protein (SP) from skeletal muscle triads. By immuno-affinity chromatography, we have purified this protein. The native protein has a molecular mass of 630-800 kD, as determined by gel filtration and rate zonal centrifugation. Within the limits of the methods used, the basic unit of the SP appears to be a dimer. In electron micrographs, it is shown to exhibit a circular profile with a diameter of approximately 100 A. In thin section analysis, the protein is frequently observed as parallel tracks of electron-dense particles bordering a translucent core. We suggest that the basic unit of the junctional structure is a dimer of 300-kD subunits and that four such entities constitute the intact SP. The purified protein has been used to develop polyclonal antibodies. By immunoelectron microscopy using immunogold probes, the SP has been localized to the junctional gap of the triad. By attaching the SP to an affinity resin, three proteins have been identified as forming associations with the SP. The Mrs of the proteins are 150, 62, and 38 kD; the 62-kD protein is calsequestrin.

Animals↗
PubMed1986

Effects of cimetidine and ranitidine on local anesthetic central nervous system toxicity in mice.

The effects of cimetidine (5-200 mg/kg), ranitidine (1-100 mg/kg), or saline on local anesthetic central nervous system toxicity was studied when administered 20 min before the administration of lidocaine (30-80 mg/kg) or 2-chloroprocaine (50-250 mg/kg) to female white CD-1 mice. All drugs were administered intraperitoneally. The 50% convulsive dose (CD50) was determined for each drug and dose combination. Pretreatment with cimetidine resulted in a significant, dose-dependent decrease in the CD50 of lidocaine, whereas ranitidine pretreatment did not significantly alter the lidocaine CD50. A dose of cimetidine (15 mg/kg) that caused a significant decrease in 2-chloroprocaine CD50 (from 180 to 110 mg/kg) caused only an insignificant decrease in lidocaine CD50 (from 59 to 54 mg/kg). If these results can be applied to clinical doses in humans, ranitidine may be a safer premedicant than cimetidine with regard to interactions with local anesthetics.

Anesthetics, Local↗
PubMed1985

Biochemical characterization of mucous glycoproteins synthesized and secreted by hamster tracheal epithelial cells in primary culture.

Hamster tracheal epithelial cells growing on type I collagen gel synthesize and secrete high molecular weight glycoconjugates which elute in the void volume upon Sepharose CL-4B column chromatography. The presence of any proteoglycans in this void volume material was ruled out based on both enzymatic analysis and behavior on DEAE-ion exchange chromatography. Based on the incorporation of radioactive precursors, followed by strong acid hydrolysis or neuraminidase digestion, the material was shown to contain sialic acid, fucose, galactose, N-acetylglucosamine, N-acetylgalactosamine, and sulfate. Complete susceptibility to papain digestion and reductive beta-elimination suggests that the material consists of O-linked glycoproteins. The identification of N-acetylgalactosaminitol in the beta-eliminated oligosaccharides confirms this notion. The molecular weight of the oligosaccharides following beta-elimination ranges from 4,000 to 15,000. We conclude that the high molecular weight glyconjugates produced by hamster tracheal epithelial cells in primary culture are mucous glycoproteins based on size, sensitivity to alkaline borohydride treatment, and monosaccharide composition. Further characterization of these mucous glycoproteins showed both size and charge microheterogeneity among molecules. Detailed structural analysis of oligosaccharides of these mucous glycoproteins is currently under way.

Animals↗
PubMed1985

Possible requirement of collagen gel substratum for production of mucin-like glycoproteins by primary rabbit tracheal epithelial cells in culture.

Primary rabbit tracheal epithelial cells growing on either plastic surface or collagen gel produce high molecular weight glycoconjugates. Biochemical characterization of these materials show they are exclusively hyaluronic acid when cells are grown on plastic surface, but a mixture of hyaluronic acid and mucin-like glycoproteins when grown on collagen gel. This research suggests that the substratum plays an important role in the maintenance or differentiation or both of mucous cells in culture.

Animals↗
PubMed1984

Pharmacokinetics, placental transfer, and neonatal effects of vecuronium and pancuronium administered during cesarean section.

Vecuronium and pancuronium were compared for placental transfer, pharmacokinetic variables, and neonatal effects during cesarean section under general anesthesia. Eighteen women underwent rapid-sequence intravenous induction using d-tubocurarine, succinylcholine, thiopental, and oxygen. Immediately after tracheal intubation, an intravenous injection of vecuronium (n = 11) or pancuronium (n = 7), 0.04 mg/kg, was given. Maternal venous blood samples were obtained before induction and at frequent intervals for 4 h after administration of vecuronium or pancuronium. Also, maternal venous and umbilical-cord arterial and venous blood samples were obtained at delivery. To describe placental transfer and maternal pharmacokinetics of the drugs, serum drug concentrations were determined using single-ion-monitoring mass spectrometry. The Apgar score and Neurologic and Adaptive Capacity Score (NACS) were used to evaluate neonatal condition. Both drugs crossed the placenta, as demonstrated by low concentrations of vecuronium (8.5-26.4 ng/ml) or pancuronium (12.2-34.2 ng/ml) found in umbilical venous blood. At delivery, the ratio of the drug concentration in umbilical venous blood to that in maternal venous blood was 0.11 +/- 0.02 for vecuronium and 0.19 +/- 0.03 for pancuronium. Vecuronium had a more rapid clearance (6.4 +/- 0.4 ml X kg-1 X min-1, mean +/- SE) and a shorter elimination half-life (36 +/- 1.8 min) than pancuronium (3.0 +/- 0.1 ml X kg-1 X min-1 and 72 +/- 6 min, respectively). No other pharmacokinetic differences were found between the drugs. Neonatal outcome was not affected adversely by either muscle relaxant, as assessed by Apgar scores and NACSs . The short duration of action, the minimal placental transfer, and the apparent lack of clinical neuromuscular effects on the newborn suggest that vecuronium should be a useful muscle relaxant for cesarean section.

Adult↗
PubMed1984

Occupational burns.

Certain occupations cause particular kinds of burn. Minor burns are usually neglected. Observation of industrial workers during routine annual medical examination found many workers with minor occupational burns and their sequalae. Some were so characteristic that they can be regarded as occupational marks.

Burns↗
PubMed1983

Yeast 2-microns plasmid DNA replication in vitro: purification of the CDC8 gene product by complementation assay.

Extracts of the yeast Saccharomyces cerevisiae support DNA replication on exogenous yeast 2-microns plasmid DNA templates. A crude extract from a S. cerevisiae cell division cycle mutant, cdc8-1, expressed the temperature-sensitive phenotype since it could be inactivated at 42 degrees C in vitro. This heat-inactivated extract was fully complemented by the addition of either wild-type or cdc8-1 single-stranded DNA binding protein (SSB). restoration by SSB of the activity of the mutant cell extract allowed replication like that of a wild-type crude extract, as shown by bidirectional DNA synthesis from the in vivo origin. The DNA binding protein specifically stimulates the reaction catalyzed by yeast DNA polymerase I, a true DNA replicase, using the hybrid of phi X174 single-stranded DNA and a restriction endonuclease fragment as a template. It also increases processivity of DNA polymerase I at least 10-fold. Escherichia coli SSB, but not T4 gene 32 protein, can substitute for yeast SSB. Both restoration of DNA synthesis in the heated mutant cell extract and stimulation of the DNA polymerase I reaction by SSB from cdc8-1 cells are inactivated at nonpermissive temperatures, suggesting that yeast SSB is the CDC8 gene product.

Carrier Proteins↗
PubMed1983

Epidural bupivacaine, chloroprocaine, or lidocaine for cesarean section--maternal and neonatal effects.

The effects of epidural anesthesia on maternal blood pressure, newborn Apgar scores, neonatal acid-base status, and the early neonatal neurobehavioral scale (ENNS) were studied in 54 pregnant women undergoing cesarean section. Maternal and neonatal blood levels of the local anesthetics were also determined. Group 1 (n = 16) received 0.75% bupivacaine, group 2 (n = 18) received 3% chloroprocaine, group 3 (n = 11) received 2% lidocaine, and group 4 (n = 9) received 2% lidocaine with 1:200,000 epinephrine. The incidence of maternal hypotension did not differ significantly among the four groups. Fetal outcome as determined by Apgar scores, acid-base status, and neurobehavioral testing was equally good in all four groups. At delivery, the fetal/maternal concentration ratio of bupivacaine was 0.27, that of lidocaine without epinephrine 0.48, and that of lidocaine with epinephrine 0.58. Chloroprocaine was detected in 12 maternal samples, in seven umbilical venous samples, and in six umbilical arterial samples. It is concluded that epidural anesthesia as administered in this study had no adverse effect on the newborn Apgar scores, cord acid-base status, or the ENNS.

Acid-Base Equilibrium↗
PubMed1982

Haloperidol causes irreversible damage to rat anterior pituitary lactotropes in culture.

Haloperidol, in concentrations in excess of 10(-5)M promotes the release of prolactin from rat anterior pituitary cells in monolayer culture. Histologic evidence revealed that 10(-4)M concentrations cause an irreversible cytotoxic effect that resulted in a loss of cell membrane integrity and a massive release of intracellular prolactin into the surrounding medium. Paradoxically, at 10(-5)M, haloperidol significantly inhibited prolactin release and promoted an accumulation of intracellular secretory granules. Lower concentrations had no significant effect. Although it is nominally classified as a dopamine receptor blocker and can (in lower concentrations) inhibit dopamine's suppression of prolactin release, haloperidol in higher concentration enhances prolactin release by a cytotoxic mechanism unrelated to dopamine receptor interaction.

Animals↗