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Biomedical subjects

K C Kim

Publications and source records attributed to K C Kim.

At least 19 recordsLinked to original sources

Thiol-linked peroxidase activity of human ceruloplasmin.

Human ceruloplasmin exhibited different antioxidant effects according to the electron donors in a metal-catalyzed oxidation system. Purified ceruloplasmin did not play a significant role in the protection of DNA strand breaks in the ascorbate/Fe3+/O2 system. However, when ascorbates were replaced with a thiol-reducing equivalent such as dithiothreitol, DNA strand breaks were significantly prevented by the same amount of ceruloplasmin. Ceruloplasmin did not catalyze the decomposition of H2O2 in the absence of reduced glutathione. On the contrary, ceruloplasmin showed a potent peroxidase ability to destroy H2O2 in the presence of reduced glutathione. In conclusion, the removal of H2O2 by human ceruloplasmin is not simply stoichiometric but thiol-dependent.

Antioxidants

Development of a competitive enzyme linked immuno sorbent assay to measure ceruloplasmin in gamma-irradiated rat serum.

The concentrations of ceruloplasmin could not be determined by comparing the oxidase activities toward p-phenylenediamine and o-dianisidine in rat whole serum with those of purified ceruloplasmin because of the different specific amine oxidase activities of ceruloplasmin purified from normal and gamma-irradiated rats and the lability caused by freezing and thawing. A competitive enzyme linked immunosorbent assay with antigens immobilized on the solid phase was developed to measure the ceruloplasmin in rat serum. The blocking materials and pH of the coating buffer had an effect on the amount of ceruloplasmin bound to the microtiter plates. The blocking materials nonspecifically reacted with the rabbit anti-rat ceruloplasmin IgG, and consequently the coating sensitivity was decreased and standard curves were not formed completely. Because rabbit anti-rat ceruloplasmin IgG did not show any cross reactivity with rat albumin and hemoglobin, these proteins did not affect the assay at a concentration of 200 micrograms/ml. This assay is useful for measuring the concentration of ceruloplasmin in normal and irradiated rat serum.

Animals

Production and characterization of monoclonal antibodies against rat tracheal mucins.

The objective of this study was to generate and characterize monoclonal antibodies against rat airway mucins. Therefore, it should serve as a useful tool in studying the regulation of airway mucins using various in vivo rat models that are currently available. As an antigen, we used a high molecular mass mucin preparation purified from the spent media of rat tracheal surface epithelial cells in primary culture. Seven monoclonal hybridomas were obtained, among which mAbRT03 showed the highest immunoreactivity against the mucin. All of the antibodies secreted by these hybridomas recognized carbohydrate epitopes but not sialic acid residues, since their immunoreactivity was completely abolished by treatment of the mucin with 20 mM periodate but not with neuraminidase. Further characterization of mAbRT03 showed that (1) it belongs to the IgM type, (2) it binds to high molecular mass mucins based on Western blot, (3) it could indirectly immunoprecipitate rat airway mucin--and, as we know, this is the first demonstration of immunoprecipitation of airway mucin with anti-mucin antibodies--(4) it binds to the luminal side of tracheal epithelium as well as some goblet cells based on immunohistochemistry, and (5) it also recognizes in vivo airway mucins from rats, but not from human nor hamsters, which have been purified from the airway lavage fluids. This is the first monoclonal antibody against rat airway mucin, which has been developed against purified rat airway mucins. Therefore, mAbRT03 should be able to serve as an invaluable tool in studying the regulation of airway mucins using various intact rat models that are currently available.

Animals

Identification and characterization of high molecular-mass mucin-like glycoproteins in the plasma membrane of airway epithelial cells.

A previous lectin binding study demonstrated the presence of high molecular-mass mucin-like glycoproteins (HMGP) on the surface of hamster tracheal surface epithelial (HTSE) secretory cells (Proc. Natl. Acad. Sci. USA 1987;84:9304). In the present study, we intended to isolate and characterize these HMGP from the plasma membrane of the primary HTSE cells and then to determine whether or not these membrane HMGP are Muc-1 mucins, a type of mucins originally discovered on the surface of some carcinomas. A subcellular fraction enriched with the plasma membrane was obtained using a sucrose density gradient centrifugation. This fraction contained high molecular-mass glycoconjugates which were excluded from Sepharose CL-4B gel. Biochemical characterization of these glycoconjugates revealed the following characteristics: (1) susceptibility to both pronase and mild alkaline treatments, but totally resistant to proteoglycan-digesting enzymes; (2) partitioning in the detergent phase of Triton X-114 and resistance to digestion by phosphatidylinositol phospholipase C or D; (3) a buoyant density of 1.5 g/ml based on CsCl density gradient centrifugation; (4) polydispersity in terms of both size and charge density; and (5) lack of immunoreactivity with an anti-Muc-1 mucin antibody. We conclude that the plasma membrane of HTSE cells at confluence contains HMGP, which seem to be the integral membrane proteins but different from Muc-1 mucins, and that these membrane HMGP appear to share some similarities with secreted mucins in terms of size and charge.

Animals

Minute chromosomal rearrangements detected prenatally by fluorescence in situ hybridization.

Initial approaches to prenatal diagnosis from fetal karyotyping involved application of standard cytogenetic techniques. However, when fetal samples, such as chorionic villus cells or amniocytes are used, small chromosome rearrangements cannot be easily identified because they lack a distinct banding pattern. We report here two cases with minute chromosome rearrangements detected prenatally by fluorescence in situ hybridization. The use of this technique allowed precise identification of fetal chromosome abnormalities, demonstrating its usefulness for characterizing conditions that would be difficult to diagnose correctly with conventional banding methods alone.

Adult

HDR syndrome (hypoparathyroidism, sensorineural deafness, renal dysplasia) associated with del(10)(p13).

A combination of hypoparathyroidism, sensorineural deafness, and renal dysplasia has been considered to be a new syndrome inherited in an autosomal dominant fashion; we name the condition "HDR syndrome." We describe a Japanese girl who has HDR syndrome associated with de novo del(10)(p13). The chromosome deletion suggests that the putative gene(s) responsible for HDR syndrome is located at a 10pter-->p13 region.

Adult

Structure and expression of the AWI 31 gene specifically induced by wounding in Arabidopsis thaliana.

Differential screening of an Arabidopsis cDNA library constructed from the plant tissues harvested 1 h after wounding resulted in the isolation of wound-inducible cDNA clones (Kim et al., 1994). The cDNA clones could be broadly classified into two groups according to the expression time of their transcripts. Nine clones from the 10 different wound-inducible cDNAs were rapidly induced, reaching a maximum level in approximately 1-1.5 h and then were progressively reduced after wounding. The cDNA clone AWI 31 showed steady accumulation of the transcripts and reached the maximum value at a later time point of 2.5 h and then started to decline. The corresponding gene of the AWI 31 in which the coding region was interrupted by an intron, had an open reading frame that predicted a protein of 386 amino acids. However, the gene product did not show any significant homology to other known proteins in the database. Northern hybridization study using the cDNA probe revealed that the gene was not regulated by other environmental stresses such as drought, high salt, low temperature, or a DPE herbicide treatment, indicating that the cDNA clone AWI 31 was specifically induced by wounding.

Amino Acid Sequence

ATP-induced mucin release from cultured airway goblet cells involves, in part, activation of protein kinase C.

Extracellular nucleotides stimulate mucin release by binding to the P2u receptor coupled to phospholipase C via G proteins (Br. J. Pharmacol. 103:1053-1056, 1991; Am. J. Respir. Cell Mol. Biol. 8:121-125, 1993). In the present study, we intended to investigate pathways downstream to the phospholipase C activation which is responsible for adenosine triphosphate (ATP)-induced mucin release in hamster tracheal epithelial cells in primary culture. We have found that: (1) Ca2+ ionophores (A23187 and ionomycin) did not affect mucin release even at 1 microM; (2) thapsigargin (10 microM), either alone or in combination with ATP (20 microM), did not enhance mucin release over its respective control group; (3) pretreatment of hamster tracheal surface epithelial (HTSE) cells with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM) (50 microM) did not inhibit ATP-induced mucin release; (4) 4beta-phorbol 12alpha-myristate 13-acetate (PMA, 1 microM) stimulated mucin release and its effect was completely blocked by protein kinase C inhibitors such as sphingosine (10 microM) and calphostin C (0.1 microM), whereas ATP-induced mucin release was blocked, only in part, by these inhibitors; (5) desensitization of protein kinase C by pretreatment with PMA inhibited the PMA-induced mucin release completely, however, ATP-induced mucin release was inhibited only partially. We conclude that mucin release by ATP does not require an increase in the intracellular Ca2+ level but involves the activation of protein kinase C. The results also suggest the presence of another mechanism separate from the phospholipase C-protein kinase C pathway for the ATP-induced mucin release.

Adenosine Triphosphate

Airway mucus.

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Humans

Airway goblet cell mucin: its structure and regulation of secretion.

Mucociliary clearance is a major function of the airway epithelium. This important function depends both on the physicochemical properties of the airway mucus and on the activity of the cilia. The former, in turn, is dependent mainly on the quality and quantity of mucous glycoproteins or mucins, which are produced by two different cell types, namely, goblet cells of the epithelium and mucous cells of the submucosal gland. Neither the structural nor the functional differences of mucins produced by these two cell types are yet known. The availability of primary airway epithelial cell culture systems, however, has made it possible to study the structure and regulation of airway goblet cells to some extent. The epithelial mucins are extremely hydrophobic and are associated with various macromolecules, the quality and quantity of which may also affect the physicochemical properties of the mucus. Secretion of epithelial mucins is stimulated by various factors, including a number of inflammatory agents. The recent progress in mucin molecular biological research will allow us to identify different mucin core proteins produced by those different cell types, and, hopefully, the differential functions of these mucins in health and disease.

Animals

Expression of MUC1 mucin gene by hamster tracheal surface epithelial cells in primary culture.

Primary hamster tracheal surface epithelial (HTSE) cells carry mucin-like glycoproteins on the apical surface which are releasable by neutrophil elastase. In some cancer cells, mucins are localized on the cell surface and have been shown to be encoded by the MUC1 mucin gene. The objectives of the present experiments were: (I) to determine if HTSE cells express MUC1 mucin gene; (2) if they do, to isolate and characterize the hamster MUC1 complementary DNA (cDNA); and (3) to examine the pattern of MUC1 mRNA expression at different stages of culture. Reverse transcriptase-polymerase chain reaction amplification of HTSE cell RNAs using degenerate primers based on homologous sequences between the human and mouse MUC1 genes revealed the presence of a cDNA (0.5 kb) which has an 88% similarity in sequence with the mouse MUC1 cDNA. Using this 0.5 kb cDNA as a probe, an HTSE cell cDNA library was screened to isolate a hamster MUC1 cDNA clone. Sequence analysis of the cDNA revealed that it encodes an integral membrane protein of 676 amino acids which consists of (1) an N-terminal signal sequence, (2) the tandem repeat domain encoding 12 repeats of 20 amino acids, and (3) the C-terminal region consisting of degenerate tandem repeats and a unique sequence containing both the transmembrane and cytoplasmic domains. The presence of seven tyrosine residues in the cytoplasmic domain suggests a potential role as a receptor. Finally, expression of MUC1 mucin gene in HTSE cells appears to be associated with differentiation of secretory cells.

Animals

Nucleotide-induced mucin release from primary hamster tracheal surface epithelial cells involves the P2u purinoceptor.

Mucin release by airway surface epithelial cells is regulated by extracellular adenosine triphosphate (ATP) via a P2 purinoceptor-mediated mechanism. The objective of the present experiment was to examine the possible involvement of uridine triphosphate (UTP) in this purinergic signal transduction pathway. Using primary hamster tracheal surface epithelial cells, ATP and UTP were compared in their abilities: 1) to displace ATP gamma S35-binding to intact cells; 2) to accumulate inositol phosphates; and 3) to stimulate mucin release. Finally, the presence of a P2u receptor message was examined. Our results showed that: 1) UTP was much less effective than ATP in displacing ATP gamma S35-binding (median inhibitory concentrations (IC50S) 240 vs 2.9 microM); 2) UTP was more potent than ATP in accumulating inositol phosphates (100 vs 43% increase at 2mM); 3) UTP was equipotent with ATP in stimulating mucin release; 4) Northern blot analysis of messenger ribonucleic acids (mRNAs) with a mouse P2u receptor complementary deoxyribonucleic acid (cDNA) probe revealed a single specific band (2.8 kb), partial sequencing of which showed a great homology with those of human or mouse P2u receptors. We conclude that, although both ATP and UTP are equipotent in stimulating mucin release, their binding kinetics to the cell surface are quite different, suggesting the presence of a common binding domain which may be responsible for the mucin release by these nucleotides. We suggest that the P2u purinoceptor is likely to be responsible for mucin release by these nucleotides, probably via activation of phospholipase C.

Adenosine Triphosphate

Anorectal dysfunction in systemic sclerosis.

This study was aimed to evaluate the anorectal dysfunction in systemic sclerosis (SSc) and propose the clinical significance of anorectal manometry in patients with SSc. Seven patients with SSc were evaluated with manometry for anorectal function and an additional 11 normal subjects were collected as a control group. The study group underwent esophageal manometry as well and the correlation between the degree of anorectal and esophageal dysfunction was evaluated. Patients showed a lower tolerance for balloon distention of the rectum than controls (minimal sensory volume and urgency volume, P < 0.05). The resting and squeezing pressure of the anal sphincter and the functional length of the anal canal showed no significant difference in these two groups. Rectoanal inhibitory reflex was absent in one (14%) and diminished in two (29%) of seven patients with SSc. SSc patients also showed abnormal esophageal manometry findings, notably decreased LES pressure and body amplitude of distal 2/3 esophagus. The comparison between manometric profiles of anorectum and esophagus showed no significant correlation by statistical analysis. In conclusion, our data could suggest that anorectal function may be impaired in patients with SSc which could reflect the involvement of the anorectum by the disease, and that anorectal manometric studies can be useful to detect such dysfunction in patients with SSc, even before clinical symptoms.

Adult

A case of paraesophageal hernia repaired by laparoscopic approach.

Paraesophageal hernia comprises only 2 approximately 5% of all hiatal hernias but is prone to incarceration and strangulation. For this reason they must be recognized and repaired as expeditiously as possible. The laparoscopic approach has already been successfully applied to the repair of the more common sliding hiatal hernia and it seems reasonable to propose that the paraesophageal hernia, provided it is not complicated, might also be repaired by the laparoscopic technique. We present here a case of paraesophageal hernia which has been successfully repaired by the laparoscopic approach. A 73-year-old female suffering from postprandial fullness in the retrosternal area was diagnosed preoperatively with paraesophageal hiatal hernia with gastroesophageal acid reflux and was submitted for laparoscopic repair. The procedure entailed reduction of the hernia, mobilization of the esophagogastric junction with crural repair and partial fundoplication. At the 9th-month follow-up, the patient had remained asymptomatic and follow-up studies revealed no evidence of hernia or acid reflux. As a result of this favorable experience with minimal morbidity, early hospital discharge, and effective control of symptoms without adverse sequalae, laparoscopic repair can be considered as the curative and minimal invasive method in the management of paraesophageal hernia.

Aged

A case of flutamide-induced acute cholestatic hepatitis--a case report.

Flutamide, an oral nonsteroidal, antiandrogenic, anilid compound which inhibits the uptake and binding of androgens to nuclear receptors in the prostate, is used with or without LH-RH analogues for treatment of patients with metastatic carcinoma of the prostate. Clinically significant hepatotoxicities such as toxic hepatitis, cholestatic hepatitis, hepatic failure, and even death have rarely been reported in the English literature, but no case has been reported in Korea. A 75-year-old man with metastatic carcinoma of the prostate had taken flutamide (750 mg/day) for 7 months and suddenly developed jaundice and general weakness. The findings of blood chemistries were compatible with cholestatic hepatitis, but ultrasonography, viral marker and auto-antibody studies did not reveal any attributable causes. Histologic examination of a sono-guided liver biopsy only disclosed centrilobular cholestasis, nuclear glycogenosis and mild sinusoidal lymphocytic infiltration. Discontinuation of flutamide resulted in an almost full recovery of the patient's liver function in 2 months. We, herein, report a case of flutamide-induced acute choestatic hepatitis with a brief review of the literature.

Acute Disease