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W K Lee

Publications and source records attributed to W K Lee.

At least 55 records · Page 3Linked to original sources
PubMed2000

Characterization of the ATP transporter in the reconstituted rough endoplasmic reticulum proteoliposomes.

Adenosine triphosphate (ATP) transporter from rat liver rough endoplasmic reticulum (RER) was solubilized and reconstituted into phosphatidylcholine liposomes. The RER proteoliposomes, resulting from optimizing some reconstitution parameters, had an apparent K(m) value of 1.5 microM and a V(max) of 286 pmol min(-1) (mg protein)(-1) and showed higher affinity for ATP and a lower V(max) value than intact RER (K(m) of 6.5 microM and V(max) of 1 nmol). ATP transport was time- and temperature-dependent, inhibited by 4, 4'-diisothiocyanostilbene-2,2'-disulfonic acid, which is known as an inhibitor of anion transporters including ATP transporter, but was not affected by atractyloside, a specific inhibitor of mitochondrial ADP/ATP carrier. The internal and external effects of various nucleotides on the ATP transport were examined. ATP transport was cis-inhibited strongly by ADP and weakly by AMP. ADP-preloaded RER proteoliposomes showed a specific increase of ATP transport activity while AMP-preloaded RER proteoliposomes did not show the enhanced overshoot peak in the ATP uptake plot. These results demonstrate the ADP/ATP antiport mechanism of ATP transport in rat liver RER.

ATP-Binding Cassette Transporters↗
PubMed2000

Cellular distribution of isozymes of protein kinase C in septal olfactory epithelium of mice.

To determine the presence of protein kinase C (PKC) isozymes in the septal olfactory epithelium of mice (mSOE), western blotting and immunohistochemistry were performed using antibodies against PKC isozymes. With the exception of PKC-betaI, all of the PKC isozymes were detected in the whole lysate of septal tissue layer and apparent molecular weights for each isoform were found. PKC-alpha, PKC-gamma and PKC-epsilon were detected in the olfactory glandular cells of the lamina propria, and PKC-betaI and PKC-betaII were located in the microvillar cells. Neither novel PKC nor atypical PKC was detected in olfactory glandular cells or microvillar cells, except for PKC-epsilon. PKC-lambda was localized in the mucous layer of the mSOE. Meanwhile, PKC-delta and PKC-xi were distributed in the receptor cells in the mSOE. These data demonstrate the isoform-specific expression of PKC in mSOE and suggest a role for the novel and atypical types of PKC in olfactory transduction.

Animals↗
PubMed2000

Cloning and characterization of the 5'-flanking region for the mouse phospholipase C-delta1 gene.

To date, little is known about the molecular mechanisms controlling the regulation of phospholipase C-delta1 (PLC-delta1) gene expression. To understand the mechanisms responsible for the regulation of PLC-delta1 gene expression, the 5'-flanking region of the mouse PLC-delta1 gene was isolated from a mouse genomic DNA library. Primer extension analysis revealed that there is a single transcriptional start site located at 127 bases upstream from the translation start codon in the mouse PLC-delta1 gene. DNA sequence analysis showed that the sequence around the transcriptional start site is very GC-rich and has no TATA or CAAT boxes. Transient expression of a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the 160-base-pair region from -622 to -462 upstream of the transcriptional start site includes a positive cis-acting element(s) for the efficient expression of the PLC-delta1 gene. Gel retardation analysis suggests that multiple transcription factors bind to separate sites on the promoter region. Based on these results, our study suggests that the minimal essential region located at -622 to +70 is fully sufficient to confer high-level transcriptional activity and contains high-affinity binding elements for multiple transcription factors.

Animals↗
PubMed2000

Parametric down conversion of X-ray photons.

Parametric down conversion of X-ray photons in diamond crystals was detected in two experiments, both using the phase-matching scheme first employed in the X-ray regime by Eisenberger & McCall [Phys. Rev. Lett. (1971), 26, 684-688]. The conversion events were detected by a combination of time-correlation spectroscopy and energy discrimination, using Si drift-chamber detectors. The time-correlation spectra give a direct comparison of the conversion rate over the accidental coincidence rate. Mechanisms for possible detection of false events and ways to cross check against them are discussed in detail.

Journal Article↗
PubMed2000

Performance limits of direct cryogenically cooled silicon monochromators - experimental results at the APS.

The successful use of cryogenically cooled silicon monochromators at third-generation synchrotron facilities is well documented. At the Advanced Photon Source (APS) it has been shown that, at 100 mA operation with the standard APS undulator A, the cryogenically cooled silicon monochromator performs very well with minimal (<2 arcsec) or no observable thermal distortions. However, to date there has not been any systematic experimental study on the performance limits of this approach. This paper presents experimental results on the performance limits of these directly cooled crystals. The results show that if the beam is limited to the size of the radiation central cone then, at the APS, the crystal will still perform well at twice the present 100 mA single 2.4 m-long 3.3 cm-period undulator heat load. However, the performance would degrade rapidly if a much larger incident white-beam size is utilized.

Journal Article↗
PubMed2000

Clinical associations of anti-endothelial cell antibodies in patients with systemic lupus erythematosus.

The aim of this study was to define the clinical associations of anti-endothelial cell antibody (AECA) in systemic lupus erythematosus (SLE) patients by measuring serum AECA titers to correlate with the disease activity and clinical manifestations. Forty-one SLE patients and 27 controls were studied. Serum samples were collected at the time of patient presentation with disease exacerbation and 4 weeks after the start of treatment. The disease activity was evaluated by the SLE Disease Activity Index (SLEDAI). AECA was detected by enzyme-linked immunosorbent assay (ELISA) methods with the surface antigen of the immortalized human microvascular endothelial cell line (HMEC-1). The mean immunoglobulin (Ig)G-AECA and IgM-AECA optical densities (ODs) were significantly higher in patients with SLE compared with controls [mean +/- standard deviation (SD), 0.32 +/- 0.15 vs 0.18 +/- 0.16 and 0.29 +/- 0.14 vs 0.21 +/- 0.09, respectively]. There was a positive correlation between IgG-AECA and the SLEDAI scores. The positivity rate of AECA in the groups with digital vasculitis, neuropsychiatric lupus, and anti-cardiolipin antibody was significant. In conclusion, AECA may be involved in the pathogenesis of SLE and was correlated with the disease activity. It was also associated with clinical manifestations such as digital vasculitis, neuropsychiatric lupus, and anti-cardiolipin antibody positivity.

Adult↗
PubMed2000

Clinical manifestations of Korean female gouty patients.

In an endeavor to analyse the clinical characteristics of female gout, we reviewed 36 women with gout. Twenty-seven (75%) developed the first symptomatic episode of gout after the onset of the menopause. The mean age at onset of gout was 54.3 years (range 15-87 years). Twenty-two patients (61%) had hypertension, 17 (47%) had renal insufficiency, 13 (36%) used diuretics and 10 (28%) were taking cyclosporine for a renal allograft. Tophaceous gout occurred in 10 patients (27%) and polyarticular involvement was seen in 16 (44%) at initial presentation. Five of nine premenopausal patients were taking cyclosporine and four had renal insufficiency. A comparison with a control group of 72 randomly selected male patients with gout showed that the female patients were frequently receiving diuretics at the time of the attack and had significantly lower mean uric acid excretion, whereas significantly more male patients showed heavy alcohol consumption and precipitating events for an acute attack compared with the female patients. There were no significant differences between the sexes for onset age, hypertension, renal insufficiency, distribution of joint involvement, tophi and mean serum uric acid concentration. The female patients in this study had a lower mean age at onset of gout than in previous studies, which was attributed to the inclusion of renal transplantation patients. Transplantation gout patients receiving cyclosporine lower the mean age at onset of female gout and this is an emerging problem in female gout.

Adolescent↗
PubMed2000

Heparinized bovine pericardium as a novel cardiovascular bioprosthesis.

A novel chemical modification of biological tissues was developed by the direct coupling heparin to bovine pericardium (BP). The heparinization involves pretreatment of BP using GA and followed by grafting heparin to BP by the reaction of residual aldehyde and amine group of heparin. BP was modified by direct coupling of heparin and the effect of heparin coupling on calcification was evaluated in vitro and in vivo. Heparinized BP was characterized by measuring shrinkage temperature, mechanical properties, digestion resistance to collagenase enzyme, in vitro cytotoxicity, and in vivo calcification. Thermal and mechanical properties showed that the durability of heparin-treated tissue increased as compared with fresh tissue and GA-treated tissue. Resistance to collagenase digestion revealed that heparin-treated tissue has greater resistance to enzyme digestion than did fresh tissue and GA-treated tissue. Heparinized tissue had shown to be non-cytotoxic, however, relatively high cytotoxicity was observed in the GA-treated tissues due to the release of GA. In vivo calcification study demonstrated much less calcium deposition on heparin-treated BP than GA-treated one. Obtained results attest to the usefulness of heparinized BP for cardiovascular bioprostheses.

Animals↗
PubMed2000

25,27-(6-Tosyl-3,9-dioxa-6-azaundecane-1,11-diyldioxy)-26,28-(3,6, 9-trioxaundecane-1,11-diyldioxy)calix[4]arene.

A new calix[4]-crowned azacrown ether, C(51)H(59)NO(11)S, consisting of four phenyl rings in a 1,3-alternate conformation was synthesized from the reaction of 25, 27-bis(5-chloro-3-oxapentyloxy)calix[4]crown-5 and p-toluenesulfonamide in the presence of Cs(2)CO(3). A crown-5 loop was attached on the two facing lower rims of the calix[4]arene and the N-tosyl azacrown group was attached on the other set of lower rims of the calix[4]arene backbone. This molecule seems to offer an inside cavity for the formation of a host-guest complex.

Journal Article↗
PubMed2000

Oxidation of ranitidine by isozymes of flavin-containing monooxygenase and cytochrome P450.

Rat and human liver microsomes oxidized ranitidine to its N-oxide (66-76%) and S-oxide (13-18%) and desmethylranitidine (12-16%). N- and S-oxidations of ranitidine were inhibited by metimazole [flavin-containing monooxygenase (FMO) inhibitor] to 96-97% and 71-85%, respectively, and desmethylation of ranitidine was inhibited by SKF525A [cytochrome P450 (CYP) inhibitor] by 71-95%. Recombinant FMO isozymes like FMO1, FMO2, FMO3 and FMO5 produced 39, 79, 2180 and 4 ranitinine N-oxide and 45, 0, 580 and 280 ranitinine S-oxide pmol x min(-1) x nmol(-1) FMO, respectively. Desmethyranitinine was not produced by recombinant FMOs. Production of desmethylranitidine by rat and human liver microsomes was inhibited by tranylcypromine, a-naphthoflavon and quinidine, which are known to inhibit CYP2C19, 1A2 and 2D6, repectively. FMO3, the major form in adult liver, produced both ranitidine N- and S-oxides at a 4 to 1 ratio. FMO1, expressed primarily in human kidney, was 55- and 13-fold less efficient than the hepatic FMO3 in producing ranitidine N- and S-oxides, respectively. FMO2 and FMO5, although expressed slightly in human liver, kidney and lung, were not efficient producers of ranitidine N- and S-oxides. Thus, urinary contents of ranitidine N-oxide can be used as the in vivo probe to determine the hepatic FMO3 activity.

Animals↗
PubMed2000

Feeder vessel laser photocoagulation of subfoveal choroidal neovascularization.

We performed laser photocoagulation for extrafoveal feeder vessels in a series of 10 eyes with subfoveal choroidal neovascular membranes (CNVMs) in which feeder vessels could be detected by means of indocyanine green angiography using scanning laser ophthalmoscope. On the follow up indocyanine green angiograms performed within one week after treatment, successful occlusion of the feeder vessels with nonperfusion of the corresponding CNVMs was observed in 8 eyes (80%). Four eyes showed improved visual acuities with resolution of the exudative manifestation. But within 3 months, recanalization of the feeder vessels or further proliferation of the CNVMs developed in 7 eyes. We confirmed the possibility of infarction and regression of the CNVM by feeder vessel photocoagulation, which could improve the visual acuity. But it was rarely possible to maintain them.

Aged↗
PubMed1999

Mechanism of silica- and titanium dioxide-induced cytotoxicity in alveolar macrophages.

Particles can cause cytotoxicity in pulmonary alveolar macrophages (AM). Several mechanisms to explain this cytotoxicity have been suggested. However, the exact mechanism of particle-induced cytotoxicity in AM remains to be established. Silica and TiO2 produced a concentration-dependent cytotoxicity as evidenced by loss of cell viability and fall in ATP levels. While silica induced a greater cytotoxicity, TiO2 produced a higher reduction in ATP levels. Silica increased the release of LDH, but TiO2 did not affect enzymatic release. TiO2 suppressed succinate-triggered oxygen consumption, whereas silica did not markedly change the effect of succinate on oxygen consumption. Polyinosinic acid (PI), a ligand of the scavenger receptor, inhibited the TiO2-induced fall in ATP content, but could not prevent the effect of silica on cellular ATP content. Data suggest that silica and TiO2 can induce cytotoxicity in AM, probably through different mechanisms.

Adenosine Triphosphate↗
PubMed1999

Rapid degradation of PrxI and PrxII induced by silica in Rat2 cells.

Peroxidases of the peroxiredoxin (Prx) family catalyze the reduction of H(2)O(2) and lipid peroxides. The effects of H(2)O(2), 12-O-tetradecanoylphorbol 13-acetate (TPA), and silica on the abundance of two cytosolic isoforms of Prx (PrxI and PrxII) were examined in Rat2 cells. TPA induces the production of reactive oxygen species (ROS) in various mammalian cell types, and silica induces the production of ROS in Rat2 cells. Whereas H(2)O(2) and TPA did not affect the concentration of PrxI or Prx II, silica triggered a rapid degradation of both Prx enzymes. Silica also induced degradation of the NF-kappaB inhibitor IkappaB-alpha. N-Acetylcysteine and diphenyleneiodonium, both of which inhibit the accumulation of intracellular ROS, each blocked silica-induced degradation of IkappaB-alpha but had no effect on that of the Prx enzymes, suggesting that ROS do not contribute to Prx proteolysis. The silica-induced degradation of Prx enzymes was also insensitive to the proteasome inhibitors MG132 and lactacystin, whereas IkappaB-alpha proteolysis was completely blocked by these inhibitors. Experiments with the Ca(2+) ionophore A23187 indicated that a Ca(2+)-dependent protease such as calpain might contribute substantially to silica-induced degradation of PrxII, but only moderately to that of PrxI. These results indicate that silica increases cellular oxidative stress not only by inducing ROS production, but also by triggering the degradation of Prx enzymes that are responsible for elimination of cellular ROS. Such aggravated oxidative stress might be important in the initial pathogenesis of silica-associated pulmonary diseases.

Animals↗
PubMed1999

Molecular cloning and expression analysis of a mouse phospholipase C-delta1.

We describe here the molecular cloning and expression analysis of mouse PLC-delta1 (mPLC-delta1), a key enzyme in cell signal transduction. A mouse brain cDNA library was screened in order to isolate the mPLC-delta1 cDNA. The mPLC-delta1 cDNA was 2660 bp in length. The predicted open reading frame encodes a protein of 756 amino acids with an estimated molecular mass of 85 kDa. The deduced amino acid sequence exhibits 96.9% and 92.7% identity with the sequence of rat and human PLC-delta1, respectively. The mPLC-delta1 mRNA was highly expressed in brain, heart, lung, and testis. We found that transcripts of mPLC-delta1 are present in almost all regions of mouse brain examined, implying that the enzyme may play a role in some fundamental cellular process in brain. In male reproductive tract, mPLC-delta1 mRNA was widely expressed in the epididymis as well as in the testis. In situ hybridization studies indicate that distribution of mPLC-delta1 mRNA in mouse testis is discrete and unique. The expression of mPLC-delta1 mRNA was defined in the periphery of each seminiferous tubule, especially in spermatogonia, which might imply that mPLC-delta1 plays a role in proliferation of spermatogonia. To the best our knowledge, this is the first report to demonstrate the high expression of mPLC-delta1 mRNA in spermatogonia of testis. Taken together, these results suggest that mPLC-delta1 may carry out fundamental roles in almost all of mouse tissues, especially in brain and specific roles in testis.

Amino Acid Sequence↗
PubMed1999

Purification and characterization of glutamate dehydrogenase as another isoprotein binding to the membrane of rough endoplasmic reticulum.

Glutamate dehydrogenase (GDH) was purified from rough endoplasmic reticulum (RER) in rat liver using anion-exchange and affinity chromatography. As GDH has been known as an enzyme that exists mainly in the matrix of mitochondria, the properties of purified GDH were compared with those of mitochondrial GDH. The GDH activity in 0. 1% Triton X-100-treated RER subcellular fraction was nearly the same as intact RER, whereas that of the mitochondrial fraction increased by 50% after the detergent treatment. In kinetic values, in addition, mitochondrial GDH had a higher K(m) value for NADP(+) than NAD(+), whereas the K(m) value for NAD(+) was higher than that for NADP(+) in the case of GDH of RER, which showed a difference in specificity to cofactors. Moreover, when two GDH isoproteins were incubated at 42 degrees C or treated with trypsin, GDH from RER was more stable against heat inactivation and less susceptible to proteolysis than mitochondrial GDH in both cases. In addition, GDH of RER had at least five amino acids different from mitochondrial GDH when sequences of N-terminal and several internal peptide fragments were analyzed. These results showed that GDH of RER is another isoprotein of GDH, of whose properties are different from those of mitochondrial GDH.

Amino Acid Sequence↗
PubMed1999

Imaging of the normal and abnormal pancreaticobiliary system with single-shot MR cholangiopancreatography: a pictorial review.

Magnetic resonance cholangiopancreatography (MRCP) is a rapidly evolving non-invasive imaging modality that produces images of the pancreatic duct and biliary tree without the need for intravenous or oral contrast. The images are equivalent to those from endoscopic retrograde cholangiopancreatography (ERCP), but the non-invasive acquisition avoids the morbidity and mortality associated with diagnostic ERCP. Magnetic resonance cholangiopancreatography is indicated in patients who require only a diagnostic ERCP, who fail an ERCP or who are unable to undergo ERCP due to altered post-surgical anatomy. Other evolving indications include triaging of patients with obstructive jaundice into percutaneous or endoscopic management drainage pathways depending on the site, length and nature of the duct obstruction, thereby potentially decreasing the number of failed or unsuccessful ERCP. Pre-operative identification of anomalous biliary anatomy and choledocholithiasis prior to laparoscopic cholecystectomy promise to modify the pre-operative and operative management of the patient in order to minimize the risk of duct injury and unnecessary intra-operative dissection and cholangiography. The advantages of the technique include its non-invasiveness, the absence of contrast administration, its relative operator independence and the ability to evaluate both sides of an obstructed duct, thereby accurately evaluating stricture morphology and length. The disadvantages of MRCP compared to ERCP include its lack of an immediate therapeutic solution to duct obstruction, procedural cost, unit availability and the inability to evaluate patients with pacemakers or ferromagnetic implants.

Bile Duct Diseases↗
PubMed1999

Invasiveness of Helicobacter pylori into human gastric mucosa.

BACKGROUND: Helicobacter pylori has generally been observed only in the gastric mucous layer or in the spaces between gastric mucus-secreting cells and not in the gastric epithelial cells or in the lamina propria. The purpose of this study is to determine whether H. pylori invades the gastric mucosa, using an immunoelectron microscopical examination of human gastric mucosa infected with H. pylori. MATERIALS AND METHODS: Five hundred gastric antral biopsy specimens were fixed in a periodate-lysin-paraformaldehyde solution, embedded in Lowicryl, sectioned, and examined with a light microscope. One hundred specimens moderately or severely infected with H. pylori were selected and were incubated with polyclonal rabbit anti-H. pylori antibody. The specimens were washed, incubated with 20 nm of colloidal gold-conjugated goat anti-rabbit IgG, stained with uranyl acetate and lead citrate, and observed with a transmission electron microscope. RESULTS: In one case, a bacterium was observed within the cytoplasm of a gastric mucus-secreting cell; in another case, a few bacteria were observed within the cytoplasm of a stromal cell in the lamina propria. The bacteria could be differentiated from degenerated intracellular organelles by gold particles attached to the bacteria. CONCLUSION: H. pylori rarely invade the lamina propria and gastric cells.

Adult↗