Biomedical subjects

Y Lee

Publications and source records attributed to Y Lee.

At least 91 records · Page 5Linked to original sources

PubMed2001

Synergistic activation of RSK correlates with c-fos induction in MO7e cells stimulated with GM-CSF plus Steel factor.

Steel factor (SLF) plus GM-CSF induces proliferative synergy in factor-dependent cell line MO7e and hematopoietic progenitor cells. We previously reported ERK1/2 involvement in this synergy, but its downstream signaling molecules are not defined. Here, we investigated activation of the 90-kDa ribosomal S6 kinase (RSK) proteins by measuring the phosphorylation status and in vitro kinase activity in MO7e cells. Both GM-CSF and SLF induced activation of RSK, and the combined stimulation with these two cytokines induced synergistic and persistent activation of RSK. RSK activity was reduced by PI3 kinase inhibitor LY294002 or MEK1 inhibitor PD98059, suggesting that the ERK as well as the PI3 kinase pathways are involved in regulation of RSK activity. Sensitivities of RSK activity to inhibitory drugs correlated well with those of c-fos gene induction. Taken together, synergistic activation of RSK may contribute, at least in part, to the synergistic induction of c-fos after combined stimulation with GM-CSF plus SLF.

Calcium-Calmodulin-Dependent Protein Kinases↗

PubMed2001

The interphase microtubule damage checkpoint defines an S-phase commitment point and does not require p21(waf-1).

Cell cycle checkpoints ensure orderly progression of events during cell division. A microtubule damage (MTD)-induced checkpoint has been described in G(1) phase of the cell cycle (G(1)MTC) for which little is known. The present study shows that the G(1)MTC is intact in activated T lymphocytes from mice with the p21(waf-1) gene deleted. However, p21(waf-1) gene deletion does affect the ratio of cells that arrest at the G(1)MTC and the spindle checkpoint after MTD. The G(1)MTC arrests T lymphocytes in G(1) prior to cdc2 up-regulation and prior to G(1) arrest by p21(waf-1). Once cells have progressed past the G(1)MTC, they are committed to chromosome replication and metaphase progression, even with extreme MTD. The G(1)MTC is also present in a human myeloid cell line deficient in p21(waf-1) gene expression. The p21-independent G(1)MTC may be important in cellular responses to MTD such as those induced by drugs used to treat cancer.

Animals↗

PubMed2001

Transcriptional and ERK1/2-dependent synergistic upregulation of p21(cip1/waf1) associated with steel factor synergy in MO7e.

Steel factor (SLF) plus GM-CSF induces proliferative synergy in myeloid progenitors and factor-dependent cell line MO7e. We previously reported that the protein level of cyclin-dependent kinase inhibitor p21(cip1/waf1) (p21) increased synergistically when MO7e cells were stimulated with SLF plus GM-CSF and that p21 induction was required for SLF synergistic responses. Here we show that this p21 induction is regulated at the transcriptional level. Based on use of a multiprobe RNase protection assay, the synergistic increase of p21 mRNA was unique among many cell cycle regulators. While STAT5A and 5B were activated after stimulation with GM-CSF alone or SLF plus GM-CSF, there was no difference in activation between the groups. p44/42 MAP kinase (ERK1/2) was synergistically activated by SLF plus GM-CSF, but SAPK/JNK and p38 MAP kinase were not. Synergistic induction of p21 was significantly decreased with a MEK1 inhibitor, suggesting that the ERK1/2 pathway is involved in the synergistic increase of p21 after GM-CSF plus SLF stimulation.

Cell Cycle↗

PubMed2001

Catalytic turnover of substrate benzylamines by the quinone-dependent plasma amine oxidase leads to H2O2-dependent inactivation: evidence for generation of a cofactor-derived benzoxazole.

Incubation of bovine plasma amine oxidase (BPAO) with benzylamine and various p-substituted analogues results in a time-dependent inactivation that is attributable to buildup of the H(2)O(2)-turnover product on the basis of protection afforded by coincubation with catalase. The mechanism of inactivation is distinct from that effected by H(2)O(2) itself, which requires higher concentrations. Solution studies using models for the 2,4,5-trihydroxyphenylalanine quinone (TPQ) cofactor reveal a loss of catalytic activity arising from oxidation of the dihydrobenzoxazole tautomer of the product Schiff base, that competes with hydrolytic release of benzaldehyde product. The resulting stable benzoxazole exhibits a characteristic absorption depending on the nature of the benzylamine p-substituent. For benzylamine itself, the model benzoxazole absorbs at 313 nm, in an area of strong absorption by the enzyme, whereas for 4-nitrobenzylamine, the absorption of the model benzoxazole is sufficiently red-shifted (at 365 nm) to be discerned above the background enzyme absorption. Inactivation of BPAO by 4-nitrobenzylamine is accompanied by loss of the resting TPQ anion absorption at 480 nm concomitant with generation of a new absorption near 360 nm. Resonance Raman spectra of the inactivated enzyme show a close correspondence with those for the model 4-nitrobenzylamine-derived benzoxazole. Substrate-dependent inactivation is also observed for the other two mammalian enzymes examined, equine plasma amine oxidase and human kidney amine oxidase. Catalase provides complete protection in these instances as well. Benzoxazole formation may constitute a common mechanism of inactivation of quinone-dependent amine oxidases by normal substrates in vitro if the product H(2)O(2) is permitted to accumulate. More importantly, the results suggest that the benzoxazole inactivation pathway may be important physiologically and may have influenced the distribution of amine oxidases and catalase in cells.

Amine Oxidase (Copper-Containing)↗

PubMed2001

Quantitative measurements of cerebral blood flow in patients with unilateral carotid artery occlusion: a PET and MR study.

Although it has been demonstrated that quantitative measures of cerebral blood flow (CBF) can be obtained with the singular value decomposition (SVD) algorithm, the extent to which quantitative CBF measurements can be utilized under pathophysiological conditions has not been systematically studied. A total of five healthy volunteers and five patients with unilateral carotid artery occlusion were studied. Only magnetic resonance (MR) images were acquired for the volunteer group while both MR and positron emission tomography (PET) images were acquired for the patient group. Assessments of CBF from normal volunteers compared favorably with values reported in the literature. However, while a linear relationship was observed for each patient when MR measured CBF was compared to that obtained from PET, this linear relationship diminished when all patients were analyzed as a group (r = 0.41). A correction factor (CF) was proposed that was equal to the ratio of the area of the venous output function (VOF) in each patient to the mean VOF obtained from the volunteer group. After globally scaling the CBF of each patient based on the experimentally derived CF, a substantial improvement was observed (a slope of 1.02 and r = 0.8 for the linear regression line) in the relationship between MR estimated CBF and those obtained from PET.

Analysis of Variance↗

PubMed2001

High-level production of heme-containing holoproteins in Escherichia coli.

The expression of recombinant protein is essential for the investigation of the functions and properties of heme-containing protein as an electron carrier. For the expression of fully active recombinant protein, conversion of the expressed apoprotein into holoprotein is the most important and difficult problem. In this study, a system was developed for the production of heme-containing protein in a pure, recombinant holoprotein form, using the bovine cytochrome b5 tryptic fragment and Escherichia coli bacterioferritin as heterologous and homologous heme-containing model proteins, respectively. This system is based on the slow synthesis of recombinant apoprotein, which can maintain the balanced consumption of amino acids between protein synthesis and heme synthesis, so that the synthesized apoprotein continues to act as a heme sink. From a 1-1 culture, 15 mg of cytochrome b5 and 40 mg of bacterioferritin were purified as pure holoprotein forms. Our expression system provides a rapid and simple method for obtaining large quantities of the active holo-form of heme-containing proteins.

Animals↗

PubMed2001

Cortical actin filaments in guard cells respond differently to abscisic acid in wild-type and abi1-1 mutant Arabidopsis.

Cortical actin filaments in guard cells of Commelina communis L. show signal-specific organization during stomatal movements [S.-O. Eun and Y. Lee (1997) Plant Physiol 115: 1491-1498; S.-O. Eun and Y. Lee (2000) Planta 210: 1014-1017]. To study the roles of actin in signal transduction, it is advantageous to use Arabidopsis thaliana (L.) Heynh., an excellent model plant with numerous well-characterized mutants. Using an immunolocalization technique, we found that actin deployments in guard cells of A. thaliana were basically identical to those in C. communis: actin proteins were assembled into radial filaments under illumination, and were disassembled by ABA. In addition, we examined actin organization in an ABA-insensitive mutant (abi1-1) to test the involvement of protein phosphatase 2C (PP2C) in the control of actin structure. A clear difference was observed after ABA treatment, namely, neither stomatal closing nor depolymerization of actin filaments was observed in guard cells of the mutant. Our results indicate that PP2C participates in ABA-induced actin changes in guard cells.

Abscisic Acid↗

PubMed2001

The emergence of erythromycin-resistant Streptococcus pyogenes in Seoul, Korea.

High frequencies of erythromycin-resistant streptococci were reported in Japan in the mid-1970s, and in Finland in the late 1980s, related to an increase in the consumption of macrolide antibiotics in these countries. The frequency of erythromycin-resistant Streptococcus pyogenes was reported to be only 2% in 1994, but we know that the susceptibility of the strains to antibiotics had not been tested routinely. We studied the resistance rates of Streptococcus pyogenes to various antibiotics in Seoul, Korea, where antibiotics could be purchased without prescription. From January through December, 1998, 92 isolates of group A streptococci were collected from inpatients and outpatients with pharyngotonsillitis or invasive streptococcal infections, from institutions in five different geographic areas of Seoul; one pediatric clinic, three university hospitals, and one general hospital. All isolates were serotyped by T-agglutination, and minimum inhibitory concentrations (MICs) were determined by agar dilution methods, according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). The most common T-serotype was T12 (44.6%), followed by T4 (19.6%). All the isolates tested were susceptible to penicillin, vancomycin, and cefotaxime. However, 38 isolates (41.3%) were resistant to erythromycin, 32 (34.8%) were resistant to clindamycin, and 48 (52.1%) were resistant to tetracycline. Twenty-seven of 41 isolates serotyped T12 and 3 of 18 isolates serotyped T28 were multiresistant to erythromycin, clindamycin, and tetracycline. Almost half of the isolates obtained from the five different areas in Seoul showed erythromycin resistance in Streptococcus pyogenes. Routine monitoring of antibiotic susceptibility tests and further extensive nationwide surveys are needed to determine the frequency and the extent of the spread of resistant strains in various geographic regions in Korea.

Adolescent↗

PubMed2001

Regulation of prolactin secretion during pregnancy and lactation.

Prolactin plays major roles in maintaining the corpora lutea of pregnancy and in the synthesis of milk during lactation. The hypothalamic mechanisms involved in these functions have been investigated. Mating leads to a surge of prolactin and programs daily surges during early pregnancy. The expression of Fos-immunoreactivity shows that mating activates several hypothalamic nuclei, particularly the arcuate nucleus and medial preoptic area. In the arcuate nucleus, mating is associated with Fos expression in beta-endorphin neurons, and infusion of naloxone blocks both mating-induced and diurnal prolactin surges. Tyrosine hydroxylase-immunoreactive dopamine neurons appear not to participate in surge generation. However, after day 10 of gestation the secretion of placental lactogens suppresses prolactin secretion via activation of dopamine neurons without involvement of beta-endorphin neurons. Intracerebroventricular implantation of placental lactogen-secreting cells will block pregnancy prolactin surges, increase Fos expression in dopamine neurons, and increase tyrosine hydroxylase activity. During lactation the mechanisms regulating dopamine and beta-endorphin neurons are further modified. In early lactation a prolactin-induced increase in tyrosine hydroxylase activity leads to negative feedback, but this effect is lost by mid-lactation. Overriding this negative feedback is the inhibitory effect that suckling has on dopaminergic activity. This may involve beta-endorphin-mediated inhibition of dopamine neurons, as naloxone causes a marked increase in tyrosine hydroxylase activity and suppression of circulating prolactin. However, removal of tonic dopamine inhibition is not sufficient to account for the high levels of prolactin attained during lactation, and additional releasing factors are probably involved. In situ hybrization histochemistry for the most recent candidate, prolactin-releasing peptide, suggests that this may involve brain stem neurons that co-localize noradrenaline. Thus, prolactin secretion during pregnancy and lactation involve complex interactions of regulatory factors and plasticity of neuronal responsiveness.

Animals↗

PubMed2001

Efficacy of chemotherapy for prostatic tuberculosis-a clinical and histologic follow-up study.

OBJECTIVES: To characterize the clinical features of prostatic tuberculosis and to evaluate the short and long-term results of antituberculous chemotherapy. METHODS: Eighteen patients (mean age 66.7 +/- 10.2 years) with prostatic tuberculosis were included in this study. The median pretreatment prostate-specific antigen (PSA) level was 2.7 ng/mL (range 0.3 to 31). The PSA level was greater than 4.0 ng/mL in 6 patients (33.3%). Eight patients (44.4%) received a triple-drug regimen of rifampin, ethambutol, and isoniazid for more than 6 months. The mean duration of chemotherapy was 7.6 months (range 6 to 12). Of the 8 patients, 3 underwent chemotherapy longer because of concurrent tuberculosis of other organs. Follow-up studies included digital rectal examination, total PSA determination, and transrectal prostate biopsy. RESULTS: Ten patients were eligible for regular follow-up. All the patients were symptom free during follow-up. The median length of follow-up was 3.4 years (range 1 to 9). The average number of follow-up transrectal prostate biopsies was 2.4 (range 2 to 3). The follow-up histologic findings showed nodular hyperplasia in 7 patients and chronic inflammatory cell infiltration in 3 patients. No acid-fast bacillus was found in any follow-up specimen. Similarly, subsequent transrectal biopsy showed no relapse after a median length of 3.4 years of follow-up. Of the 6 patients with elevated PSA levels, the post-treatment PSA returned to normal in 3 patients. CONCLUSIONS: Our results suggest that a triple-drug regimen of 6 months' duration can successfully control prostatic tuberculosis. Histologic follow-up is a good method for monitoring the efficacy of treatment. Transrectal prostate biopsy is an important tool for the diagnosis and follow-up of prostatic tuberculosis.

Adult↗

PubMed2001

Comparative genomic hybridization analysis reveals 3q gain resulting in genetic alteration in 3q in advanced oral squamous cell carcinoma.

We analyzed DNA sequence copy number aberrations (DSCNAs) in 17 primary oral squamous cell carcinomas (OSCCs) by comparative genomic hybridization. DSCNAs were detected frequently at 3q25-qter (7/17), Xp21 (5/17), and Xq12-q23 and 8q23-q24 (4/17), and losses were detected frequently at 13q21-q22 (5/17), 3p21-pter, 4p15-pter and 17p13 (4/17), and 8p22-pter and 9p21-pter (3/17). Four tumors showed amplifications of seven loci: 3q11-qter, 3q13, 3q26, 7q21-q22, 8q23-qter, 9p22-pter, and 12p11. The total number of DSCNAs was significantly greater in stage III and stage IV tumors than in stage I and stage II tumors (P=.008). Furthermore, 3q gain was detected preferentially in stage III and stage IV tumors (6/8) rather than in stage I and stage II tumors (1/9, P=.013). In our study, all tumors with gain of 3q also contained one or more loss(es) in common regions. On the other hand, all tumors with gain of 9p did not contain 3q gains. These observations indicate that gain of 3q and accumulation of DSCNAs are strongly associated with tumor progression in OSCC. Furthermore, 3q gain and loss of one or more additional loci in common aberration regions appears to be a group of DSCNs associated with dominant genetic pathways of leading to advanced OSCCs.

Adult↗

PubMed2001

The feasibility and safety of fluoroscopy-guided percutaneous intramyocardial gene injection in porcine heart.

BACKGROUND: Catheter-based transendocardial gene injection would be useful for the delivery of genes into the heart. We examined the feasibility and safety of percutaneous intramyocardial gene injections with fluoroscopic guidance alone. METHODS: We performed the procedure through an 8F arterial sheath inserted into the left carotid artery. In protocol 1, a mixture of India ink and normal saline was injected through a needle injection catheter in six pigs. We monitored blood pressure and ECG continuously during the procedure. Echocardiography, left ventriculography, and coronary angiography were performed. All pigs were sacrificed 2 days later and hearts were harvested. In protocol 2, a mixture of India ink and plasmid encoding CAT gene was injected in the same manner in eight pigs. Myocardial tissue was obtained 7 days after the procedure to assess gene expression. In protocol 3, four pigs were intentionally needle-perforated in the ventricular wall and were observed for 7 days. RESULTS: In protocol 1, there was no significant hemodynamic changes or serious arrhythmias during the procedure. Echocardiography and angiography revealed no evidence indicating pericardial effusion or wall motion abnormalities. Harvested hearts revealed one intramyocardial hematoma in a total of 36 injection sites. In protocol 2, the gene expression could be identified in 39 sites out of 48 injections after 7 days. In protocol 3, no animal showed signs indicating cardiac tamponade during the observation period. CONCLUSIONS: Our data suggest that fluoroscopy-guided percutaneous intramyocardial gene injection is a feasible and safe procedure, with no indication of associated significant hemodynamic changes, arrhythmias, or mortality.

Administration, Cutaneous↗

PubMed2001

Liquid chromatographic-tandem mass spectrometric urine assay for a highly metabolized cyclic ureidobenzenesulfonamide: issues concerning assay specificity and quality control preparation.

An LC-MS-MS method was validated for the quantitation of a beta(3) agonist (A) in human urine to support Phase I studies. A was designed to accelerate metabolism for weight reduction. During assay development a significant loss of A was apparent from frozen urine quality control samples. The addition of 0.75% bovine serum albumin (BSA) in urine (v/v) was required to maximize the recovery of A from urine. Urine samples were basified and extracted into methyl t-butyl ether-isopropyl alcohol (90:10, v/v). The organic layer was washed, evaporated, reconstituted, and injected onto a 5 cm, C8 HPLC column prior to MS-MS analysis. The standard curve was linear from 5 to 500 ng/ml. Intraday precision for peak area ratios from BSA urine samples at seven separate concentrations over a range of 5-500 ng/ml (n=5) was <4.0% and calculated concentrations were within 91-115% of nominal concentrations. Interday precision for BSA urine quality control (QC) samples at four separate concentrations (n=10 of each) was <5.0% and individual calculated concentrations were within 90-111% of nominal concentrations. This work emphasizes that potential metabolites and quality control standards should be prepared and assayed as early as possible in method development, especially before the sample collection section of the clinical protocol is prepared. The methods described here have wide utility to other compounds containing basic benzene sulfonamides and to beta3 agonist candidates.

Adrenergic beta-Agonists↗

PubMed2001

Trauma experience of North Korean refugees in China.

OBJECTIVES: We examined trauma experience and mental health conditions among North Korean migrants in China. METHODS: Personal interviews of 170 North Korean "food refugees" in China were conducted in 1999. Structured questionnaires were used to identify specific trauma experience and symptoms of post-traumatic stress, anxiety, and depression. RESULTS: All participants surveyed reported trauma exposure, with food and water shortage (93%) and illness without access to medical care (89%) being the most frequently cited events. Post-traumatic stress disorder was suspected in 56% of the respondents. Above-threshold scores on anxiety and depression were found in 90% and 81% of the cases, respectively. CONCLUSIONS: The burden on the part of North Korean migrants in China who experience trauma appears to be very high. An international call for action is warranted to monitor and safeguard the mental health status of this vulnerable segment of people and respond to their urgent psychiatric and medical care needs.

Adult↗

PubMed2001

Simultaneous capture of metal, sulfur and chlorine by sorbents during fluidized bed incineration.

Metal capture experiments were carried out in an atmospheric fluidized bed incinerator to investigate the effect of sulfur and chlorine on metal capture efficiency and the potential for simultaneous capture of metal, sulfur and chlorine by sorbents. In addition to experimental investigation, the effect of sulfur and chlorine on the metal capture process was also theoretically investigated through performing equilibrium calculations based on the minimization of system free energy. The observed results have indicated that, in general, the existence of sulfur and chlorine enhances the efficiency of metal capture especially at low to medium combustion temperatures. The capture mechanisms appear to include particulate scrubbing and chemisorption depending on the type of sorbents. Among the three sorbents tested, calcined limestone is capable of capturing all the three air pollutants simultaneously. The results also indicate that a mixture of the three sorbents, in general, captures more metals than a single sorbent during the process. In addition, the existence of sulfur and chlorine apparently enhances the metal capture process.

Adsorption↗

PubMed2001

Expansins: ever-expanding numbers and functions.

Expansins were first identified as cell-wall-loosening proteins that, at least in part, mediate pH-dependent extension of the plant cell wall and growth of the cell. More recently, it has been realized that expansins belong to two protein families, the alpha-and beta-expansins, and that they appear to be involved in regulating, besides cell expansion, a variety of plant processes, including morphogenesis, softening of fruits, and growth of the pollen tube of grasses through the stigma and the style. The Arabidopsis genome contains 26 alpha-expansin genes and the rice genome at least 26. There are more beta-expansin genes in monocots than in dicots, at least 14 in rice and five in Arabidopsis. Expansin genes are differentially regulated by environmental and hormonal signals, and hormonal regulatory elements have been found in their promoter regions. An analysis of exon/intron structure led to the hypothesis that alpha-and beta-expansins evolved from a common ancestral gene.

Arabidopsis↗

PubMed2001

cDNA cloning of two isoforms of ornithine carbamoyltransferase from Canavalia lineata leaves and the effect of site-directed mutagenesis of the carbamoyl phosphate binding site.

The immunoscreening method was used to isolate cDNAs of 1323 bp (ClOCT1) and 1433 bp (ClOCT2) encoding two ornithine carbamoyltransferases (OCT, EC 2.1.3.3) from the cDNA expression library of Canavalia lineata leaves constructed in a lambdaZAP Express vector. ClOCT1 and ClOCT2 encode 359 and 369 amino acids, respectively. The N-terminals of deduced amino acid sequences of the two cDNAs showed typical features of the transit peptide of chloroplast targeting proteins. The ornithine-binding domain (FMHCLP) and catalytic domain (HPXQ) of ClOCT1 and ClOCT2 and the carbamoyl phosphate (CP)-binding site of ClOCT1 (SMRTR) are identical to OCTs of other plant species, pea and Arabidopsis thaliana. However, the CP-binding site sequence of ClOCT2, SLRTH, has not yet been reported. Both ClOCT1 and ClOCT2 cDNAs were expressed in Escherichia coli BL21 (DE3) by using expression vector pET30a. Recombinant ClOCT1 protein showed 14 times higher ornithine-dependent OCT activity than canaline-dependent OCT activity. In contrast, recombinant ClOCT2 protein showed 13 times higher canaline-dependent OCT activity than ornithine-dependent OCT activity. The two amino acids of the CP-binding site of ClOCT2 (SLRTH) were combinatorially changed to those of the CP-binding site of ClOCT1 (SMRTR) by site-directed mutagenesis. When Leu-118 of ClOCT2 was changed to Met, ornithine-dependent activity was increased significantly. It is assumed that the substrate specificity of ClOCT1 or ClOCT2 proteins partially depends on the amino acid sequence of the CP-binding site.

Amino Acid Sequence↗

PubMed2001

Optimal operational factors for nitrite accumulation in batch reactors.

The environmental factors that affected the accumulation of nitrite in nitrifying reactors were investigated using a mixed culture. A batch reactor with 50 mg-N/l of ammonia was used. The pH, temperature and dissolved oxygen concentration were varied. The concentration of unionized free ammonia also changed with the oxidation of ammonia and the variation of pH and temperature. The accumulation of nitrite was affected sensitively by pH and temperature. A higher nitrite concentration was observed at pH 8-9 or temperature around 30 degrees C. The dissolved oxygen also affected, giving the highest nitrite accumulation at around 1.5 mg/l. These were the favored conditions for nitrite production. The free ammonia concentration influenced the nitrite accumulation also, by inhibiting nitrite oxidation. The inhibition became apparent at a concentration of approximately 4 mg/l or above, but insignificant at below 1 mg/l. Thus, simultaneously high free ammonia concentration and maximum specific ammonia-oxidation rate (above 15 x 10(-3) mg-N/mg-VSS x h) were needed for a significant nitrite accumulation. When the two conditions were met, then the highest accumulation was observed when the ratio of the maximum specific oxidation rate of ammonia to the maximum specific oxidation rate of nitrite (ka/kn) was highest. Under the optimal operating conditions of pH 8, 30 degrees C and 1.5 mg/l of dissolved oxygen, as much as 77% of the removed ammonia accumulated in nitrite.

Algorithms↗