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Biomedical subjects

C H Kim

Publications and source records attributed to C H Kim.

At least 289 records · Page 16Linked to original sources
PubMed1996

An open clinical trial of fluoxetine in the treatment of premature ejaculation.

There have been an increased number of recent reports on orgasm-related sexual dysfunction coincident with selective serotonin reuptake inhibitor (SSRI) treatment. In contrast, it has also been reported that SSRIs improve sexual dysfunction. Low doses of clomipramine and paroxetine, potent 5-hydroxytryptamine reuptake blockers, have been found to retard ejaculation time. We hypothesized that the SSRI fluoxetine might be effective in treating premature ejaculation. In an 8-week open-label clinical study, 11 male patients with premature ejaculation were treated with fluoxetine. After a washout period of 2 weeks, each patient was assigned to receive fluoxetine, 20 mg/day for 2 weeks, and then titrated to 60 mg/day, depending on the patient's tolerability and clinical response. A within-subjects comparison of pre- and posttreatment intravaginal ejaculation latency time revealed a significant improvement. Fluoxetine treatment produced significant improvements in self-visual analogue scale scores for sexual desire, anxiety for rapid ejaculation, and partner's satisfaction with ejaculation and overall sexual function. These data suggest that serotonergic antidepressants may be effective in treating rapid ejaculation in men and underline the need to carry out a double-blind, placebo-controlled trial to confirm these results.

Adult↗
PubMed1996

A novel autonomously replicating sequence (ARS) for multiple integration in the yeast Hansenula polymorpha DL-1.

Several autonomously replicating sequences of Hansenula polymorpha DL-1 (HARSs) with the characteristics of tandem integration were cloned by an enrichment procedure and analyzed for their functional elements to elucidate the mechanism of multiple integration in tandem repeats. All plasmids harboring newly cloned HARSs showed a high frequency of transformation and were maintained episomally before stabilization. After stabilization, the transforming DNA was stably integrated into the chromosome. HARS36 was selected for its high efficiency of transformation and tendency for integration. Several tandemly repeated copies of the transforming plasmid containing HARS36 (pCE36) integrated into the vicinity of the chromosomal end. Bal 31 digestion of the total DNA from the integrants followed by Southern blotting generated progressive shortening of the hybridization signal, indicating the telomeric localization of the transforming plasmids on the chromosome. The minimum region of HARS36 required for its HARS activity was analyzed by deletion analyses. Three important regions, A, B, and C, for episomal replication and integration were detected. Analysis of the DNA sequences of regions A and B required for the episomal replication revealed that region A contained several AT-rich sequences that showed sequence homology with the ARS core consensus sequence of Saccharomyces cerevisiae. Region B contained two directly repeated sequences which were predicted to form a bent DNA structure. Deletion of the AT-rich core in region A resulted in a complete loss of ARS activity, and deletion of the repeated sequences in region B greatly reduced the stability of the transforming plasmid and resulted in retarded cell growth. Region C was required for the facilitated chromosomal integration of transforming plasmids.

Base Sequence↗
PubMed1996

Mechanism of relaxations to C-type natriuretic peptide in veins.

C-type natriuretic peptide (CNP) is an endothelium-derived peptide that shares structural homology with atrial natriuretic peptide (ANP). CNP causes greater endothelium-independent relaxations in veins compared with arteries. Relaxations to CNP in porcine coronary arteries are mediated by hyperpolarization of the smooth muscle membrane. Experiments were designed to investigate the mechanism(s) by which CNP causes relaxation in canine femoral veins. Rings of canine femoral veins without endothelium were suspended for measurement of isometric force in organ chambers. Concentration-response curves to CNP were obtained in veins contracted with either endothelin-1 (10(-8) M), KCl (40 mM), phenylephrine (10(-6) M) or prostaglandin F2 alpha (2 x 10(-6) M) in the absence and presence of BQ-123 (10(-6) M), NG-monomenthyl-L-arginine (L-NMMA; 10(-4) M), HS-142-1 (10(-5) M), methylene blue (10(-5) M), or potassium channel blockers, tetraethylammonium chloride (TEA; 10(-3) M), charybdotoxin (10(-7) M), glibenclamide (10(-7) M), or apamin (10(-7) M). Relaxations to CNP were significantly attenuated when the tissue was contracted with KCl and endothelin-1. During contraction to either phenylephrine or prostaglandin F2 alpha, relaxations to CNP were inhibited by HS-142-1, methylene blue, TEA, and charybdotoxin, but not by L-NMMA, glibenclamide, or apamin. In separate experiments, guanosine 3',5'-cyclic monophosphate increased twofold within 10-60 s after the addition of CNP (10(-8) M). These data suggest that CNP mediates relaxation of canine femoral veins through activation of large-conduction, calcium-activated potassium channels and activation of particulate and soluble guanylate cyclase.

Animals↗
PubMed1996

Retinoic acid signaling is required during early chick limb development.

In the chick limb bud, the zone of polarizing activity controls limb patterning along the anteroposterior and proximodistal axes. Since retinoic acid can induce ectopic polarizing activity, we examined whether this molecule plays a role in the establishment of the endogenous zone of polarizing activity. Grafts of wing bud mesenchyme treated with physiologic doses of retinoic acid had weak polarizing activity but inclusion of a retinoic acid-exposed apical ectodermal ridge or of prospective wing bud ectoderm evoked strong polarizing activity. Likewise, polarizing activity of prospective wing mesenchyme was markedly enhanced by co-grafting either a retinoic acid-exposed apical ectodermal ridge or ectoderm from the wing region. This equivalence of ectoderm-mesenchyme interactions required for the establishment of polarizing activity in retinoic acid-treated wing buds and in prospective wing tissue, suggests a role of retinoic acid in the establishment of the zone of polarizing activity. We found that prospective wing bud tissue is a high-point of retinoic acid synthesis. Furthermore, retinoid receptor-specific antagonists blocked limb morphogenesis and down-regulated a polarizing signal, sonic hedgehog. Limb agenesis was reversed when antagonist-exposed wing buds were treated with retinoic acid. Our results demonstrate a role of retinoic acid in the establishment of the endogenous zone of polarizing activity.

Animals↗
PubMed1996

[Localization and characteristics of lactate and malate dehydrogenase in the sparganum and adult worm of Spirometra erinacei].

This study investigated the enzyme histochemical localization and characteristics of lactate (LDH) and malate dehydrogenase (MDH) related with the oxidation-reduction metabolism in the sparganum and adult of S. erinacei. By enzyme histochemical assay, activity of LDH was strong in the tegument and subtegumental muscle layers of the adult and sparganum. Activity of MDH was strong in the tegument of the sparganum and subtegumental muscle layers of the adult. However it was weak in the tegument of the adult. By electrophoresis, 45 kDa band was major and common in LDH of adults and spargana. The 150 kDa molecule was the major and common band in MDH of adults and r-spargana (from experimentally infected rats). By isoelectrofocusing, isoelectric points (PI) of 4 MDH isozyme from adult worm were 6.0, 6.5, 6.7 and 7.1, respectively. PI 6.0 was the major band. The active range of pH for MDH was about pH 6 approximately 8 and the optimum pH was pH 7. The effective temperature on the MDH was about 30 degrees C approximately 50 degrees C and the optimum temperature was about 40 degrees C in spargana and adult worm. In the stability against heat, when MDH was heated at 85 degrees C for 10 seconds, the activity was denatured perfectly. Maximum activity of MDH was 19.4 unit in the s-sparganum (from snakes), 24.5 unit in the r-sparganum (from rats) and 108.0 unit in the adult worm. The maximum activity was higher in adults than in spargana. The present result showed us that the nutrients absorbed through the tegument were transferred into inner tissues and were utilized as the source of metabolism. According to the habitat of the parasite, the isozymes of LDH and MDH are activated differently, and by this different activation the sparganum and adult can adapt themselves to parasitic circumstances.

Animals↗
PubMed1996

[Characteristics of alkaline and acid phosphatase in Spirometra erinacei].

This study was done to investigate the enzyme-histochemical localization and characteristics of alkaline and acid phosphatase related with metabolism in sparganum and adult of Spirometra erinacei. By the enzyme-histochemical assay, the alkaline and acid phosphatases were localized in the tegument and subtegumental musculature of sparganum and adult, but not in the parenchyma. The activities of alkaline phosphatase were stronger in the tegument than in the subtegumental musculature, and activities of acid phosphatase were stronger in the tegument of adults than those of sparganum. The 2 isozymes of alkaline and acid phosphatases were separated from s-sparganum (from snake) and r-sparganum (from experimentally infected rats) respectively, but 4 isozymes of Alp and 3 isozymes of Acp were separated from adult worms by electrophoresis. In isozyme Alp, the 66 kDa was the common isozyme, but 130 kDa isozyme of Acp was the common isozyme in spargana and adult worms. By isoelectrofocusing, 4 isozymes (PI 7.9, 7.7, 6.5 and 6.3) and 2 isozymes (PI 7.9 and 7.7) of alkaline phosphatase were separated from adults and spargana, respectively. In the stability against heat, activity of alkaline phosphatase was denatured perfectly after heating at 90 degrees C for 40 seconds. The optimum pH and temperature for activity of alkaline phosphatase were about pH 10 and 50 degrees C, respectively. The maximum activity (unit) of alkaline phosphatase was 22.0 in s-sparganum, 25.0 in r-sparganum and 215.0 in adult worms, so that the maximum activity was revealed higher in adults than spargana. As the result from above, we observed that alkaline and acid phosphatases were functioned mainly in the tegument and subtegumental musculature, and the isozymes of phosphatase were activated differently according to habitat of the parasites. The spargana and adult worms carry out the parasitism by adapting themselves to parasitic circumstance with these enzymes.

Acid Phosphatase↗
PubMed1996

Variation in artificial heart acceleration and sound production with prosthetic valve selection in vitro.

In an attempt to explore methods to reduce total artificial heart (TAH) acceleration and sound production, in vitro measurements of TAH acceleration and sound were made when using a variety of prosthetic valves in a test ventricle. A miniature, uniaxial, high fidelity accelerometer was glued to the housing of a UTAH-100 left ventricle adjacent to the inflow and outflow ports and parallel to the axis of diaphragm excursion. A miniature, high fidelity contact microphone was glued to the opposite side of the ventricular housing between the inflow and outflow ports. Data was collected over a range of heart rates, ventricular filling volumes and control modes while using tilting disc valves (TDV) and polymer trileaflet valves (PTV). For both valve types, the peak systolic acceleration impulse was lower (approximately equal to 50%) when the ventricle was fully rather than partially filled and the peak diastolic acceleration impulse was lower (approximately equal to 50%) when the ventricle was fully rather than partially ejected. The magnitude of the acceleration with PTVs was approximately equal to 20x less than the TDVs (0.5 to 2.2 g vs. 10 to 49g). The magnitude of the sound production was also considerably less with the PTVs (28 to 49 db vs. 50 to 64 db). Diastolic acceleration and sound production was approximately twice the systolic value for the TDVs; the reserse was true for PTV sound production. These data demonstrate the substantial reduction in TAH impulse acceleration and sound production by selecting PTVs over TDSs. With PTVs, the TAH impulse acceleration is the same as the natural heart. Consequently, this ability to lower TAH acceleration and sound production to the level of the natural heart may lead to a reduction in component wear, patient discomfort and other undesirable consequences of TAH implantation.

Aorta↗
PubMed1996

Site-specific crosslinks of yeast U6 snRNA to the pre-mRNA near the 5' splice site.

We have introduced a single photochemical crosslinking reagent into specific sites in the central domain of U6 to identify the sites that are in close proximity to the pre-mRNA substrate. Four distinct U6 snRNAs were synthesized with a single 4-thiouridine (4-thioU) at positions 46, 51, 54, and 57, respectively. Synthetic U6 RNA containing the 4-thioU modifications can functionally reconstitute splicing activity in cell-free yeast splicing extracts depleted of endogenous U6 snRNA. Upon photoactivation with UV (>300 nm), 4-thioU at position 46 forms crosslinks to pre-mRNA near the 5' splice site at nt +4, +5, +6, and +7 in the intron, whereas 4-thioU at position 51 crosslinks to the pre-mRNA at positions -2, -1, +1, +2, +3, and at the invariant G in the lariat intermediate. All crosslinks are dependent on the presence of ATP and the splicing substrate. The two crosslinks to the pre-mRNA from position 46 and 51 of U6 can also occur in prp2 heat-inactivated yeast splicing extracts blocked immediately prior to the first chemical step. Significantly, the crosslink from position 51 can undergo subsequent splicing when the mutant extract is complemented with functional Prp2 protein in a chase experiment, indicating that the crosslink reflects a functional interaction that is maintained during the first step. The crosslink to lariat intermediate appears when the mutant spliceosomes are complemented with functional Prp2 protein added exogenously. This experiment is a paradigm for future studies in which different mutant extracts are used to establish the stage in assembly at which particular RNA-RNA interactions defined by unique crosslinks occur.

Actins↗
PubMed1995

Expression and functional analysis of a novel isoform of gicerin, an immunoglobulin superfamily cell adhesion molecule.

We have cloned a novel cDNA of gicerin, a cell adhesion molecule belonging to the immunoglobulin superfamily. Both gicerin isoforms share the same extracellular domain, which has five immunoglobulin-like loop structures and a transmembrane domain as s-gicerin, but differ in the cytoplasmic tail domain. As the newly identified form has a larger cytoplasmic domain than the previously reported form, we refer to them as l-gicerin and s-gicerin, respectively. l-gicerin is transcribed from a distinct mRNA containing an inserted sequence not found in s-gicerin mRNA which caused a frameshift for the coding region for a cytoplasmic domain. Previous studies demonstrated that gicerin showed a doublet band of 82 and 90 kDa in chicken gizzard smooth muscle. We report that the 82-kDa protein corresponds to s-gicerin and the 90-kDa protein to l-gicerin. We also found that the two gicerin isoforms are expressed differentially in the developing nervous system. Functional analysis of these gicerin isoforms in stable transfectants revealed that they had differ in their homophilic adhesion properties, as well as in heterophilic cell adhesion assayed with neurite outgrowth factor. In addition, these isoforms have neurite-promoting activity by their homophilic adhesion, but differ in their ability to promote neurite outgrowth.

Amino Acid Sequence↗
PubMed1995

Rapid turnover and impaired cell-surface expression of the human folate receptor in mouse L(tk-) fibroblasts, a cell line defective in glycosylphosphatidylinositol tail synthesis.

The human folate receptor (hFR) is a plasma membrane protein that is anchored to the membrane via a glycosylphosphatidylinositol (GPI) tail in some cell types. The KB hFR cDNA sequence predicts a hydrophobic, alpha-helical 31-residue carboxyl terminus that is thought to be the signal for cleavage and attachment of the GPI tail. Alternatively, this region may serve as a transmembrane domain if GPI attachment is not efficient. In this study, we investigated the latter possibility by expressing the hFR in L(tk-) cells, cells that are unable to synthesize GPI tails for attachment to membrane proteins. We also transfected the same hFR cDNA into Chinese hamster ovary (CHO) cells, cells that can anchor proteins by either a GPI tail or a transmembrane domain. Neither parental cell line expresses detectable levels of folate receptor as determined by folic acid binding assays, Western analysis, or Northern analysis. In L(tk-) cells, we found that the recombinant hFR is not expressed on the cell surface, but is rapidly degraded (t1/2 < or = 4 h). Most (> 95%) of the recombinant hFR remains Endo H sensitive, suggesting retention in the endoplasmic reticulum. In contrast, transfected CHO cells express functional hFR protein at the cell surface, the half-life of the hFR is long (t1/2 > or = 24 h), and the Endo H glycosylation pattern of the recombinant hFR is consistent with normal processing through the Golgi apparatus. Therefore, in the absence of a GPI tail, the hFR is not sorted to the cell surface and the incompletely processed hFR protein is unstable.

Animals↗
PubMed1995

Determination of a necdin cis-acting element required for neuron specific expression by using zebra fish.

To determine cis-acting elements required for neuron specific expression of a necdin gene, we tried to use zebra fish assay system in vivo instead of cell lines in vitro. Various expression vectors carrying upstream sequences of necdin gene fused to MEKA (lacZ) gene as a reporter were injected into fertilized zebra fish embryos and then the expression of the reporter gene was analyzed by the whole mount immunochemical method. No promoter activity was obtained with a construct carrying sequence from -63 to +63 of the necdin gene, while promoter activity with preferential skin expression was obtained with a construct having sequence from -86 to +28. Further upstream sequence from -173 to +28 exhibited neuron specific expression as well as that from -845 to +63. These results indicate that a cis-acting element responsible for neuron specific expression is located in an 87bp sequence from -173 to -87 of necdin gene.

Animals↗
PubMed1995

Role of the MoFe protein alpha-subunit histidine-195 residue in FeMo-cofactor binding and nitrogenase catalysis.

Site-directed mutagenesis and gene-replacement procedures were used to isolate mutant strains of Azotobacter vinelandii that produce altered MoFe proteins in which the alpha-subunit residue-195 position, normally occupied by a histidine residue, was individually substituted by a variety of other amino acids. Structural studies have revealed that this histidine residue is associated with the FeMo-cofactor binding domain and probably provides an NH-->S hydrogen bond to a central bridging sulfide located within FeMo-cofactor. Substitution by a glutamine residue results in an altered MoFe protein that binds but does not reduce N2, the physiological substrate. Although N2 is not a substrate for the altered MoFe protein, it is a potent inhibitor of both acetylene and proton reduction, both of which are otherwise effectively reduced by the altered MoFe protein. This result provides evidence that N2 inhibits proton and acetylene reduction by simple occupancy of a common active site. N2 also uncouples MgATP from proton reduction catalyzed by the altered MoFe protein but does so without lowering the overall rate of MgATP hydrolysis. Thus, the quasi-unidirectional flow of electrons from the Fe protein to the MoFe protein that occurs during nitrogenase turnover is controlled, in part, by the substrate serving as an effective electron sink. Substitution of the alpha-histidine-195 residue by glutamine also imparts to the altered MoFe protein hypersensitivity of both its acetylene reduction and N2 binding to inhibition by CO, indicating that the imidazole group of the alpha-histidine-195 residue might protect an Fe contained within the FeMo-cofactor from attack by CO. Finally, comparisons of the catalytic and spectroscopic properties of altered MoFe proteins produced by various mutant strains suggest that the alpha-histidine-195 residue has a structural role, which serves to keep FeMo-cofactor attached to the MoFe protein and to correctly position FeMo-cofactor within the polypeptide matrix, such that N2 binding is accommodated.

Azotobacter vinelandii↗
PubMed1995

Electron spin echo envelope modulation spectroscopic analysis of altered nitrogenase MoFe proteins from Azotobacter vinelandii.

Electron spin echo envelope modulation (ESEEM) spectroscopy was used to study changes in the polypeptide environment of the FeMo-cofactor that were elicited by amino-acid substitutions within the nitrogenase MoFe protein alpha-subunit. A previous ESEEM study [Thomann et al. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 6620] detected modulation arising from nitrogen coupled to the S = 3/2 spin system of the FeMo-cofactor (Fe7S9Mo:homocitrate). Such modulation was found to be sensitive to the substitution of alpha-195His by alpha-195Asn as indicated by whole-cell ESEEM analysis of mutant strains from Azotobacter vinelandii. Subsequent structural studies revealed that the alpha-195His residue does not provide direct N-coordination to the cluster but is within hydrogen-bonding distance of one of a set of three sulfides that bridge the FeMo-cofactor subcluster fragments. In the present work, the ESEEM analysis is extended to both partially purified alpha-195Asn MoFe protein and purified MoFe protein from an additional mutant strain in which alpha-195His is replaced by alpha-195Gln. The dramatic decrease in the intensity of the ESEEM signal resulting from the alpha-195Asn substitution in whole cells was confirmed for the case of the isolated alpha-195Asn MoFe protein. In contrast, substitution of alpha-195His by alpha-195Gln caused no detectable change in the modulation. Simulations of the alpha-195His and alpha-195Gln ESEEM data give quadrupole parameters of e2qQ = 2.2 MHz and eta = 0.5.(ABSTRACT TRUNCATED AT 250 WORDS)

Azotobacter vinelandii↗
PubMed1995

Characterization of a DNA fragment carrying the raw starch-digesting alpha-amylase and salt-dependent alpha-amylase genes from Bacillus circulans F-2.

A 5.4 kb HindIII DNA fragment carrying the gene encoding raw starch-digesting alpha-amylase (RSDA), has been previously cloned from Bacillus circulans F-2 and expressed in Escherichia coli [Kim et al. (1990) Biochim. Biophys. Acta 1048, 2233-2238]. Interestingly, when the cell extract of E. coli harboring a plasmid carrying this fragment was incubated with 1 M NaCl, it exhibited about 10 times higher enzyme activity than when assayed without NaCl. Differential zymograms showed two different amylase activities: one for RSDA and the other for a salt-dependent alpha-amylase (SDA). Even though RSDA activity was detected without NaCl, SDA activity was detected only in high concentrations of NaCl. SDA activity was fully detected at above 1 M NaCl. Results from subcloning of the genes, fractionation analysis of cell extracts, and immunological assays clearly suggested that the two amylases are genetically distinct and that genes for both enzymes are closely linked on the 5.4 kb DNA fragment.

Bacillus↗
PubMed1995

Substrate specificity and detailed characterization of a bifunctional amylase-pullulanase enzyme from Bacillus circulans F-2 having two different active sites on one polypeptide.

Bacillus circulans F-2 amylase-pullulanase enzyme (APE) displayed dual activity with respect to glycosidic bond cleavage. The enzyme was active on alpha-1,6 bonds in pullulan, amylopectin, and glycogen, while it showed alpha-1,4 activity against malto-oligosaccharides, amylose, amylopectin, and soluble starch, but not pullulan. Kinetic analysis of the purified enzyme in a system which contained both pullulan and amylose as two competing substrates was used to distinguish the dual specificity of the enzyme from the single-substrate specificity known for pullulanases and alpha-amylases. Enzyme activities were inhibited by some metal ions, and by metal-chelating agents with a different mode. The enzyme-inhibitory results of amylase and pullulanase with Hg2+ and Co2+ ions were different, indicating that the activation mechanisms of both enzyme activities are different. Cyclomaltoheptaose inhibited both alpha-amylase and pullulanase activities with inhibition constants (Ki) of 0.029 and 0.06 mg/ml, respectively. Modification with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide confirmed a carboxy group at the active sites of both enzymes. The N-terminal sequence of the enzyme was: Ala-Asp-Ala-Lys-Lys-Thr-Pro- Gln-Gln-Gln-Phe- Asp-Ala-Leu-Trp-Ala-Ala-Gly-Ile-Val-Thr-Gly-Thr-Pro-Asp-Gly-Phe. The purified enzyme displayed Michaelis constant (Km) values of 0.55 mg/ml for amylose, and 0.71 mg/ml for pullulan. When both amylose and pullulan were simultaneously present, the observed rate of product formation closely fitted a kinetic model in which the two substrates are hydrolyzed at different active sites. These results suggest that amylopullulanases, which possess both alpha-1,6 and alpha-1,4 cleavage activities at the same active site, should be distinguished from APEs, which contain both activities at different active sites on the same polypeptide. Also, it is proposed that the Enzyme Commission use the term 'amylase-pullulanase enzyme' to refer to enzymes which act on starch and cleave both alpha-1,6-bonds in pullulan and alpha-1,4 bonds in amylose at different active sites.

Amino Acid Sequence↗