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Biomedical subjects

K J Lee

Publications and source records attributed to K J Lee.

At least 19 recordsLinked to original sources
PubMed1992

Analysis of antiepileptic drugs in human plasma using micellar electrokinetic capillary chromatography.

We describe a method for the simultaneous determination of antiepileptic drugs (ethosuccimide, phenytoin, primidone, phenobarbital, carbamazepine and valproic acid) by micellar electrokinetic capillary chromatography using sodium dodecyl sulphate as the micellar phase. Factors affecting the micellar electrokinetic separation were studied for the quantitative determination of these drugs in human plasma. The confirmation of the peaks and the specificity of the method were investigated by combining multiwavelength detection with micellar electrokinetic capillary chromatography.

Anticonvulsants↗
PubMed1992

Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene.

To examine the relationship between the cardiac and skeletal muscle gene programs, the current study employs the regulatory (phosphorylatable) myosin light chain (MLC-2) as a model system. Northern blotting, primer extension, and RNase protection studies documented the high level expression of the cardiac MLC-2 mRNA in both mouse cardiac and slow skeletal muscle (soleus). Transgenic mouse lines harboring a 2100- or a 250-base pair rat cardiac MLC-2 promoter/luciferase fusion gene were generated, demonstrating high levels of luciferase activity in cardiac muscle, and only background luminescence in slow skeletal muscle and non-muscle tissues. As assessed by in situ hybridization, immunofluorescence, and luminescence assays of luciferase reporter activity in various regions of the heart, both the endogenous MLC-2 gene and the MLC-2 luciferase fusion gene were expressed exclusively in the ventricular compartment, with expression in the atrium at background levels. Point mutations within the conserved regulatory sites HF-1a and HF-1b significantly cripple ventricular muscle specificity, while mutation of the single E-box site was without effect, suggesting that ventricular muscle-specific expression occurs through an E-box-independent pathway. This study provides direct evidence that the cis regulatory sequences in the cardiac/slow twitch MLC-2 gene which confer cardiac and skeletal muscle-specific expression can be clearly segregated, suggesting that distinct regulatory programs may have evolved to control the tissue-specific expression of this single contractile protein gene in cardiac and skeletal muscle.

Amino Acid Sequence↗
PubMed1992

Separation of theophylline and its analogues by micellar electrokinetic chromatography: application to the determination of theophylline in human plasma.

Micellar electrokinetic chromatographic separation of theophylline and its analogues was investigated using sodium dodecyl sulphate (SDS) as a micellar phase. The effects of pH, micelle concentration, applied voltage and temperature on the separation and preliminary quantitative analysis were studied for the determination of theophylline in human plasma. The data indicate that this technique could be used as the reference or routine method of theophylline measurement in therapeutic drug monitoring.

Chromatography↗
PubMed1992

Transactivation of human immunodeficiency virus type 1 long terminal repeat-directed gene expression by the human foamy virus bel1 protein requires a specific DNA sequence.

Human foamy virus (HFV) encodes the transcriptional transactivator bel1. The bel1 protein transactivates HFV long terminal repeat (LTR)-directed gene expression by recognizing a region in U3. It also transactivates human immunodeficiency virus type 1 (HIV-1) LTR-directed gene expression in transient transfection assays. To identify the specific region in HIV-1 LTR responsible for bel1 action, we examined the effect of bel1 on chloramphenicol acetyltransferase (CAT) gene expression in transfected cells with a series of mutant HIV-1 LTR/CAT plasmids. The region between -158 and -118 from the transcription initiation site, immediately upstream of the core enhancer element, was identified as responsible for the transactivation by bel1. In addition, bel1 transactivated a heterologous promoter when this region was positioned upstream of it in the sense and antisense orientations. Optimal transactivation of the HIV-1 LTR by bel1 did not require an intact TAR sequence, suggesting that the binding of tat to the TAR sequence is not a prerequisite for bel1 function in HIV-1 LTR-directed gene expression. In the region of the HIV-1 LTR that is necessary for the bel1-mediated transactivation, we have found a sequence which is conserved between HIV-1 and HFV. Our results suggest that the bel1 action on HIV-1 seems to be mediated by a specific DNA sequence which is shared by both the HIV-1 LTR and HFV LTR.

Base Sequence↗
PubMed1992

Management of retinal detachment associated with choroidal coloboma using perfluoroperhydrophenanthrene (Vitreon).

Vitreous surgery using an internal tamponading agent and chorioretinal adhesion is commonly used to repair retinal detachment associated with choroidal coloboma. The detachment may, however, recur if fluid continues to leak through a retinal defect in the area of the coloboma, despite internal tamponade. We present a case in which a recurrent retinal detachment associated with coloboma following vitreous surgery with SF6 was successfully repaired in outpatient surgery using perfluoroperhydrophenanthrene (Vitreon) and endolaser photocoagulation.

Aged↗
PubMed1992

Hemoglobin adduct and hepatic- and urinary bladder-DNA adduct levels in rapid and slow acetylator Syrian inbred hamsters administered 2-aminofluorene.

The levels of covalently bound arylamine-hemoglobin and DNA adduct formation were used as dosimeters to measure the effect of acetylator genotype and sex on the metabolic conversion of the carcinogen, 2-aminofluorene, to reactive intermediates. A single high dose of 2-aminofluorene (60 mg/kg b.wt. i.p.) was administered to male and female homozygous rapid (Patr/Patr) acetylator hamsters (MHA/SsLaK) and homozygous slow (Pats/Pats) acetylator hamsters (Bio. 82.73/H). By using 32P-postlabeling assay methodology, a sole nonacetylated DNA adduct, which cochromatographed with authentic N-(deoxyguanosin-8-yl)-2-aminofluorene was detected at 3, 6, 12, 18 or 24 hr postdosing in liver and urinary bladder DNA of both rapid and slow acetylator hamsters. The highest levels were detected at 18 hr post 2-aminofluorene injection at which time the average levels of hepatic 2-aminofluorene-DNA adducts were similar between male and female rapid and slow acetylators. By comparison, the levels of 2-aminofluorene-DNA adducts in the urinary bladder at 18 hr were about 4-fold lower than in the liver, and were significantly greater in homozygous rapid than in homozygous slow acetylator counterparts (P less than .01). In both the liver and urinary bladder, the levels of 2-aminofluorene-DNA adducts were independent of sex. In contrast to the DNA adduct data, the levels of 2-aminofluorene-hemoglobin adducts, evaluated by capillary gas chromatography-mass spectrometry, were significantly higher in the homozygous slow acetylators than in homozygous rapid acetylators. However, there again were no differences between males and females.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylation↗
PubMed1991

Free solution capillary electrophoresis of proteins using untreated fused-silica capillaries.

Numerous efforts have been made to separate proteins by capillary zone electrophoresis (CZE). The most common optimization techniques are changing the pH of the running buffer, coating the capillary surface with a hydrophilic polymer, or using additives in the sample solution. Surface coatings and solution additives can reduce the adsorption of the protein onto the capillary surface, but they diminish the separation efficiency and the resolution of CZE. This paper reports the successful separation of proteins in a untreated fused-silica capillary by raising the pH of the running buffer and washing between runs with 1.0 M sodium hydroxide. Under these conditions, model proteins and proteins in human serum have been determined by CZE. It is shown that the results from CZE are compatible with those of sodium dodecyl sulphate-polyacrylamide gel electrophoresis.

Blood Proteins↗
PubMed1991

Protocol for resolving protein mixtures in capillary zone electrophoresis.

The separation of protein mixtures by capillary zone electrophoresis can be plagued by wall adsorption of the protein components, causing peak broadening and distortion. A method is presented for overcoming this problem by adding ethylene glycol to the protein sample and by choosing the running buffer and protein sample to be at different pH values and molarities. This protocol appears to work for a wide class of proteins having different molecular weights and pI values. The method has been applied to the analysis of proteins in human serum. Compared to the traditional method of agarose gel electrophoresis, the present method is more rapid and offers better resolution, suggesting its potential as a clinical diagnostic of certain disease states.

Blood Proteins↗
PubMed1991

Isolation and characterization of the disconnected gene of Drosophila melanogaster.

Mutations in the disco (disconnected) gene prevent the establishment of stable connections between the larval optic nerves, the Bolwig's nerves, and their target cells in the brain during embryonic development. The failure of this initial connection is associated with aberrant development of the optic lobes which are largely degenerate in the mutant adult fly. In order to understand the role of disco in establishing this connection, we isolated and characterized the disco gene. A 22 kb DNA fragment can completely rescue the mutant phenotype. A single transcript, 2.9 kb in length, is found in this region and is expressed throughout development of the fly. We determined the nucleotide sequence of the disco gene to be unique when compared with sequences in a number of databases. The predicted amino acid sequence contains a region with similarity to the consensus established for the zinc finger motif. Mobilization of a P-element inserted near the gene resulted in the deletion of the 5' end of the gene and produced flies indistinguishable from those carrying the disco allele.

Amino Acid Sequence↗
PubMed1991

Expression of the disconnected gene during development of Drosophila melanogaster.

Proper development of the larval visual nerve, Bolwig's nerve, of Drosophila melanogaster requires the wild type function of the disconnected (disco) gene. In disco mutants, the nerve does not make stable connections with its targets in the larval brain. We have begun to explore the role of disco in the formation of the nervous system by examining the distribution of disco mRNA and protein in embryos and third instar larvae using in situ hybridization and antibody staining respectively. No differences between the distribution patterns of the two products are detected; disco is expressed in many tissues including both neural and non-neural cells. Many of the cells which express disco undergo extensive movement during development as they participate in major morphogenetic movements. Antibody staining shows that the protein is found in the cell nucleus. Products of the disco gene are detected in cells near the terminus of the growing Bolwig's nerve. In embryos homozygous for either of two mutant alleles of disco, the disco protein is absent near the nerve terminus, although protein distribution elsewhere is indistinguishable from wild type.

Animals↗
PubMed1991

Clinical and immunologic evaluations of reactive dye-exposed workers.

To evaluate type 1 hypersensitivity to reactive dyes, its prevalence, and its relationship to respiratory dysfunction, we studied clinical and immunologic features, including skin prick tests. RAST, and bronchoprovocation tests, of 309 employees working in a reactive-dye industry. Our survey revealed that 78 (25.2%) employees had work-related lower respiratory symptoms associated with or without nasal, skin, or eye symptoms. Among these employees, 38 (48.7%) had nonspecific bronchial reactivity. Thirteen demonstrated immediate (6), dual (6), or late only (1) asthmatic responses after inhalation of four kinds of reactive-dye solutions. Twenty-five employees demonstrated immediate skin responses to black GR dye, and 21 reacted to orange 3R. Fifty-three employees (17%) had specific serum IgE antibody against black GR and orange 3R-human serum albumin conjugate. Specific IgE was detected more frequently in symptomatic employees (30%) and smokers (100%). No association was found between atopy and specific IgE binding. The RAST-inhibition tests of black GR revealed significant inhibitions by black GR-human serum albumin conjugate and minimal inhibitions by unconjugated black GR. Orange 3R RAST-inhibition tests revealed significant inhibitions by conjugated forms of black GR and orange 3R and some inhibitions by two unconjugated dyes, suggesting an immunologic cross-reactivity between these dyes. These findings suggested that reactive dyes could induce immunologic responses, most likely IgE-mediated.

Adult↗
PubMed1991

Relationship between threonine dehydratase and biosynthesis of tylosin in Streptomyces fradiae.

To elucidate the repression mechanism of ammonium ions on the biosynthesis of tylosin in Streptomyces fradiae NRRL 2702, enzyme activities involved in the metabolism of the aspartate family of amino acids were evaluated in relation to the ammonium ion concentration and tylosin production. It was found that aspartate aminotransferase was essential for both cell growth and tylosin production. However, both threonine dehydratase and valine dehydrogenase were repressed by supplemented ammonium ions at concentrations higher than 50 mM. Threonine dehydratase was purified from cell-free extracts by acetone precipitation, ion-exchange chromatography and gel filtration, and its molecular mass was estimated to be 67,200 Da. The optimum pH and temperature for threonine dehydratase activity were 7.5 and 25 degrees C, respectively, and the Km value for threonine under these optimum conditions was 21 mM. The inhibition pattern of ammonium ions on the activity of threonine dehydratase appeared to be a mixed type.

Ammonia↗
PubMed1991

Influence of amineptine on changes of blood pressure evoked by norepinephrine and dopamine.

The influence of amineptine, an antidepressant currently employed having mainly selective dopaminergic neurochemical activity, on the pressor responses evoked by norepinephrine (NE) and dopamine (DA) was studied in anesthetized whole rats. Amineptine at doses of 0.5, 1.5, and 5.0 mg/kg/30 min infused into the femoral vein of the rat caused a dose-related inhibition of the pressor responses of NE and DA. The hypertensive responses of NE and DA augmented by pretreatment with reserpine, a catecholamine depletor, were also clearly depressed following the infusion of amineptine with a rate of 1.5 mg/kg/30 min. Furthermore, the pressor responses of NE and DA potentiated by pretreatment with debrisoquin, a sympathetic neuron blocker, were markedly diminished after pretreatment with the infusion of amineptine at the above same rate (1.5 mg/kg/30 min). These experimental results demonstrate that amineptine causes an inhibitory effect on the pressor responses evoked by NE and DA. It is thought that the amineptine effect may be due to the blockade of the peripheral adrenergic alpha-receptors in addition to the previously described uptake inhibition of dopamine.

Animals↗
PubMed1991

Fine structure and sugar transport functions of the tegument in Clinostomum marginatum (Digenea: Clinostomatidae): environmental effects on the adult phenotype.

Digenean flukes can be classified into 3 groups according to their location in the host: the lumen of the alimentary canal or associated organ, body cavity or tissue, and external surfaces. We obtained adults of Clinostomum marginatum that had matured in these 3 habitats and compared the fine structure and glucose transporting capacity of their teguments. Adults from the esophagus of herons, Ardea herodias, had thick, smooth teguments and took up glucose by facilitated diffusion, the type of transport that is Na(+)-independent and insensitive to phlorizin. By contrast, the surfaces of adults cultured from metacercariae in body cavities of laboratory mice were amplified 3-5-fold due to numerous irregular projections of the tegument. Glucose transport by these worms was largely Na(+)-dependent and inhibited by phlorizin, indicating active transport. Ectoparasites from herons' mouths had relatively thick, smooth teguments, but these worms always were encrusted with bacteria and yeast that are known to absorb and metabolize glucose. Most of the attached bacteria, and the apparent glucose uptake associated with their presence, were removed by treating the worms with antibiotics prior to transport assays. As facilitated diffusion and active transport are operational simultaneously in metacercariae, the type of transport function, if any, expressed in the adult is determined by environmental conditions associated with the worm's habitat.

Animals↗