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D S Lee

Publications and source records attributed to D S Lee.

At least 19 recordsLinked to original sources
PubMed1992

Catalytic properties of the cloned amylase from Bacillus licheniformis.

A gene encoding a new amylolytic enzyme of Bacillus licheniformis (BLMA) has been cloned, and we characterized the enzyme expressed in Escherichia coli. The genomic DNA of B. licheniformis was double-digested with EcoRI and BamHI and ligated the pBR322. The transformed E. coli was selected by its amylolytic activity, which carries the recombinant plasmid pIJ322 containing a 3.5-kilobase fragment of B. licheniformis DNA. The purified enzyme encoded by pIJ322 was capable of hydrolyzing pullulan and cyclodextrin as well as starch. It was active over a pH range of 6-8 and its optimum temperature was 50 degrees C. The molecular weight of the enzyme was 64,000, and the isoelectric point was 5.4. It degraded soluble starch by cleaving maltose units preferentially but did not attack alpha-1,6-linkage. The enzyme also hydrolyzed pullulan to panose units exclusively. In the presence of glucose, however, it transferred the panosyl moiety to glucose with the formation of alpha-1,6-linkage. The specificity of transferring activity is evident from the result of the maltosyl-transferring reaction which produces isopanose from maltotriose and glucose. The molecular structure of the enzyme deduced from the nucleotide sequence of the clone maintains limited similarity in the conserved regions to the other amylolytic enzymes.

Amino Acid Sequence↗
PubMed1992

Proteolytic modification of raw-starch-digesting amylase from Bacillus circulans F-2 with subtilisin: separation of the substrate-hydrolytic domain and the raw substrate-adsorbable domain.

Raw starch-digesting amylase (BF-2A, 93,000 Da) from Bacillus circulans F-2 was converted into two components during digestion with subtilisin. The two components were separated and designated BF-2A' (63 kDa) and BF-2B (30 kDa), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in Km, Vmax and in their specific activity for soluble starch hydrolysis. However, its absorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was differed greatly from that of BF-2A. The stability of the enzymes decreased below pH 5.5 and at 50 degrees C, while it was quite stable even at pH 12. On the other hand, the smaller peptide (BF-2B) could be adsorbed onto raw starch. From these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption and also contributes to the original enzyme-to-enzyme stabilization. A proposed model of the raw-starch-digesting enzyme from this strain is extensively discussed.

Amylases↗
PubMed1991

Absent or faint renal uptake on bone scan. Etiology and significance in metastatic bone disease.

A review of 14,296 unselected bone scans identified 889 scans showing absent or faint renal uptake. The majority of cases were associated with renal insufficiency (816/889; 91.8%), while widespread metastatic bone disease was the most common cause in a group of patients without renal disease (53/889; 6.0%). Of the 140 patients with prostate cancer, 108 (77.1%) had evidence of bone metastasis, 19 of whom (17.6%) revealed absent or faint renal uptake, demonstrating that poor renal uptake is more frequently associated with prostate cancer than with any other malignancy. Of note was that 162 out of 328 (49.4%) patients with stomach cancer at varying stages showed evidence of bone metastasis, and 14 of them (8.6%) showed poor renal images on bone scan. Interestingly, ankylosing spondylitis and rheumatoid arthritis were occasional causes of lack of renal activity (4 and 3 cases, respectively). A case of adult-form osteopetrosis, showing strikingly increased uptake mainly in the long bones with markedly diminished renal uptake, was also included in this study. Of the 53 bone scans with metastatic disease showing poor renal uptake, 44 (83.0%) revealed evidence of diffuse or multiple metastases in both spine and ribs, while 49 (92.5%) showed malignant involvement in three or more regions and 35 (66.0%) in four or more regions, suggesting widespread bone involvement in most cases.

Adult↗
PubMed1990

Stimulation of prostaglandin synthesis by hyperoxia in perinatal rat lung cells.

Prostaglandins (PGs) have been implicated in the development of pulmonary oxygen toxicity. We tested the hypothesis that hyperoxia modulates PG synthesis in a differentiation-arrested primary lung cell culture model in the rat at three developmental ages: day-20 gestation (term = 22 days), days 1 and 3 after birth. The time courses of the response to hyperoxia were defined in preconfluent lung cells as well as in growth-arrested, confluent cells. From days 4-8 after plating in growth medium containing 10% carbonstripped fetal bovine serum, exposure to 95% O2, in contrast to 1% O2, inhibited cell proliferation but significantly enhanced the production of PGI2 and, to a lesser extent, PGE2 at all three ages. The capacity to metabolize exogenous arachidonic acid (AA) to PGI2 was also increased two-to threefold (P less than 0.01). Cellular release of lactate dehydrogenase, a measure of O2 toxicity, remained unchanged during exposure to 1% O2 but increased fivefold between 48 and 96 h after exposure to hyperoxia (from 2% total to 10.5%, P less than 0.01). In confluent, growtharrested cells, under serum-free conditions, exposure to hyperoxia for 24-48 h resulted in a similar induction of PG synthesis. Our results suggest that hyperoxia stimulates PG synthesis in the perinatal rat lung and that this effect is independent of cell growth or the presence of serum. We speculate that this hyperoxia-induced PG synthesis is a relatively early response to oxidant stress and may serve as an useful early marker for O2 toxicity in perinatal lung cells.

Animals↗
PubMed1990

Localization of In-111 antimyosin Fab and 99mTc-pyrophosphate in reperfusion myocardial infarction model.

The myocardial uptake of In-111 antimyosin Fab and Tc-99m pyrophosphate was studied in dogs undergoing coronary artery occlusion for 90 minutes followed by reperfusion. The regional myocardial blood flow was determined by injecting Sc-46 labeled microsphere and was related to the relative concentrations of In-111 antimyosin and Tc-99m pyrophosphate. There was an inverse linear correlation between In-111 antimyosin Fab localization and the regional blood flow in both the subendocardial (r = 0.81) and subepicardial myocardium (r = -0.80). The greatest uptake of antimyosin was observed in areas of severe blood flow reduction (0-10% of normal). On the other hand, there was no correlation between the Tc-99m pyrophosphate uptake and the degree of blood flow reduction. Maximal subendocardial localization of Tc-99m degree of blood flow reduction. Maximal subendocardial localization of Tc-99m pyrophosphate was observed in areas where the blood flow was reduced to 31-50% of the normal. In the case of the subepicardium, the greatest uptake was localized to areas of 0 to 10% of the normal flow. In addition, there was significant myocardial uptake in regions where the blood flow was minimally reduced (greater than 81%). This study suggests that In-111 antimyosin Fab is a specific and quantitative tool in the evaluation of myocardial necrosis.

Animals↗
PubMed1989

Effects of reactive oxygen species on prostacyclin production in perinatal rat lung cells.

A differentiation-arrested primary cell culture model was used to examine the role of reactive oxygen species in the control of prostacyclin (PGI2) production in the perinatal rat lung. Coincubation of the lung cells with arachidonic acid (AA) and xanthine (X, 0.25 mM) plus xanthine oxidase (XO, 10 mU/ml) or with AA and glucose (25 mM) plus glucose oxidase (25 mU/ml) augmented the AA-induced PGI2 output. Superoxide dismutase (10 U/ml) did not alter the X + XO effect, whereas catalase (10 U/ml) eliminated both X + XO and glucose plus glucose oxidase effects. H2O2 (1-200 microM) showed a dose-related biphasic augmentation with peak stimulation at 20 microM. Catalase again blocked this effect, but dimethylthiourea, a hydroxyl radical scavenger, did not. A 20-min pretreatment of the cells with X + XO, glucose plus glucose oxidase, or H2O2, however, diminished the capacity of the cells to convert exogenous AA to PGI2. This pretreatment effect was also blocked by catalase. The responses were similar in lung cells obtained from day 20 rat fetuses (term = 22 days) and 1-day-old newborn rats. Lactate dehydrogenase release was not detected during treatment periods but increased significantly after exposure to reactive oxygen species.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Ketoprostaglandin F1 alpha↗
PubMed1989

Decrease in plasma prostaglandin E2 is not essential for the establishment of continuous breathing at birth in sheep.

Depression of prostanoid concentrations by indomethacin induces continuous breathing in fetal sheep, but it is not known whether this is associated with changes in fetal behaviour. Furthermore, the relationship between changes in prostaglandin E2 (PGE2) concentration after delivery and the appearance of continuous breathing has not been examined. We hypothesized that the decrease in fetal PGE2 by infusion of indomethacin would induce continuous breathing and a change in behaviour such that the fetus should come to resemble a newborn lamb; and coinciding with the establishment of continuous breathing at birth, PGE2 concentrations would decrease to a critical level below that present in the fetus. We found that continuous breathing in fetal sheep induced by infusion of indomethacin was related to a decrease in PGE2 from 436 +/- 114 to 189 +/- 73 pg/ml (P less than 0.005) but that this was not associated with fetal wakefulness. In addition, measurements of carotid arterial PGE2 concentrations showed that the beginning of continuous breathing after birth occurred at a plasma concentration of PGE2 of 1245 +/- 260 pg/ml, a value about three times higher than the 422 +/- 53 pg/ml measured in the fetus during breathing activity. Together these findings suggest that PGE2 is not primarily involved in the establishment of continuous breathing at birth.

Animals↗
PubMed1988

Search for the optimal sequence of the ribosome binding site by random oligonucleotide-directed mutagenesis.

Synthetic DNA duplexes corresponding to the ribosome binding site (RBS) were synthesized through the phosphite method on solid support. The synthetic RBS DNA with partial random sequences was inserted into an appropriate site between the lpp-lac promoter and the beta-galactosidase structural gene in plasmid pMKT2. The level of beta-galactosidase expression was correlated with the color intensity of the recombinant colonies on X-gal plates. The bluest colonies were isolated and characterized with respect to beta-galactosidase enzyme activity and RBS sequence. There was good correlation between color intensity and the level of the enzyme activity, and this provided a reliable phenotypic screening method in the search for the optimal regulatory sequences. Novel RBS sequences obtained here show not only the unique nucleotide distribution, but also strong complemetarity to the 3' end region of 16S rRNA, from which could be deduced a generalized RBS sequence, the position of the SD region, and the 16S rRNA position mediated during translation initiation.

Bacterial Proteins↗
PubMed1988

Shivering following retrobulbar block.

A case of a severe but brief period of shivering following a retrobulbar block (RBB) is presented. The shivering occurred within two minutes after completion of the RBB and subsided gradually within five minutes, without specific treatment. The patient remained conscious during the episode of shivering. The shivering was so abrupt and severe as to be misjudged as a seizure, but its onset appeared to be slower than a seizure. The mechanism of shivering appeared to be the central spread of local anaesthetic solution into the brain stem, along the optic nerve. Shivering may be a warning sign of brain stem anaesthesia and demands special care to anticipate life-threatening complications.

Anesthetics, Local↗
PubMed1988

Effect of morphine on breathing and behavior in fetal sheep.

To define the dose response of apnea and breathing to morphine we studied 12 fetuses at 116-141 days of gestation using our window technique. We instrumented the fetus to record electrocortical activity (ECoG), eye movements (EOG), diaphragmatic activity (integral of EMGdi), heart rate, carotid blood pressure, and amniotic pressure. Saline and morphine in doses of 0.03, 0.1, 0.5, 1, and 3 mg/kg were injected in random order in the jugular vein of the fetus during low-voltage ECoG. Fetuses were videotaped for evaluation of fetal behavior. We found 1) that saline did not elicit a response; 2) apnea, associated with a change from low- to high-voltage ECoG, increased from 2.2 +/- 1.5 (SE) min in two fetuses at a dose of 0.03 mg to 20 +/- 6.3 min in seven fetuses at 3 mg/kg (P less than 0.005); 3) the length of the breathing responses, associated with a change from high- to low-voltage ECoG, were 15 +/- 1.8 and 135.9 +/- 18.1 min (P less than 0.0005); 4) integral of EMGdi X frequency, an index equivalent to minute ventilation, increased from 1,763 +/- 317 arbitrary units to 10,658 +/- 1,843 at 1.0 mg/kg and then decreased to 7,997 +/- 1,335 at 3.0 mg/kg. These changes were related to a steady increase in integral of EMGdi, whereas frequency decreased at 3 mg/kg. There was an increase in breathing response to morphine plasma concentrations or morphine doses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Ventilatory response to hypoxia in unanesthetized newborn kittens.

We studied the ventilatory response to hypoxia in 11 unanesthetized newborn kittens (n = 54) between 2 and 36 days of age by use of a flow-through system. During quiet sleep, with a decrease in inspired O2 fraction from 21 to 10%, minute ventilation increased from 0.828 +/- 0.029 to 1.166 +/- 0.047 l.min-1.kg-1 (P less than 0.001) and then decreased to 0.929 +/- 0.043 by 10 min of hypoxia. The late decrease in ventilation during hypoxia was related to a decrease in tidal volume (P less than 0.001). Respiratory frequency increased from 47 +/- 1 to 56 +/- 2 breaths/min, and integrated diaphragmatic activity increased from 14.9 +/- 0.9 to 20.2 +/- 1.4 arbitrary units; both remained elevated during hypoxia (P less than 0.001). Younger kittens (less than 10 days) had a greater decrease in ventilation than older kittens. These results suggest that the late decrease in ventilation during hypoxia in the newborn kitten is not central but is due to a peripheral mechanism located in the lungs or respiratory pump and affecting tidal volume primarily. We speculate that either pulmonary bronchoconstriction or mechanical uncoupling of diaphragm and chest wall may be involved.

Aging↗
PubMed1988

Histiocytic medullary reticulosis: radiologic diagnosis of splenic infarction--a case report.

A case of histiocytic medullary reticulosis with splenic infaraction from a 23-year-old male is presented. Radiologic findings on selective spleen scintigraphy and abdominal CT are described. Selective spleen scintigraphy showed huge, multilobulated spleen with numerous photon-deficient areas in it and peripherally. Abdominal CT showed large peripheral band-like low density and infiltrative lesion in spleen with accompanying intraabdominal lymphadenopathy. Histoligical features were consistent with HMR in spleen and liver specimens.

Adult↗
PubMed1987

HLA and Graves' disease in Koreans.

The HLA-A, B, C and DR antigen distribution in 128 Korean patients with Graves' disease was compared with that in 220 controls. The frequency of HLA-B13, DR5 and DRw8 (relative risk 3.8, 4.4, and 2.3, respectively) was significantly increased in patients with Graves' disease. There was no significant correlation between the presence of these HLA antigens and the clinical features.

Female↗
PubMed1985

Malignant hyperthermia: a possible new variant.

A young healthy male, who had three consecutive episodes of postoperative hyperthermia was anaesthetized with special precautions to prevent malignant hyperthermia. Despite neuroleptic anaesthesia and dantrolene pretreatment, the patient experienced post-anaesthetic hyperthermia. The patient's clinical picture was almost identical to the symptoms experienced by two of his maternal relatives. All three experienced nausea, vomiting, muscle cramps and high fever which occurred between five to seven hours after general anaesthesia. The serum potassium (K) and creatinine phosphokinase (CPK) levels determined during the hyperthermic episode and on the next day were not elevated. On the basis of the patient's family history, his clinical picture, and his laboratory data, we speculate that this patient might have a form of malignant hyperthermia or a possible new variant.

Adolescent↗
PubMed1985

Human serum vitamin B12 assay methods--a review.

The clinical importance of a reliable human serum vitamin B12 assay to aid the diagnosis of pernicious anemia (PA) cannot be overemphasized. Our review of the literature indicates that a reference method for the quantitation of serum vitamin B12 (serum B12) with the required accuracy, precision and rapidity has not been reported to-date. Controversies, debates and criticisms over human serum B12 assays (especially commercial kits) have been common-place. Various methods of quantitation of B12 reviewed in this communication include: microbiological, radioisotopic, high-performance liquid chromatography (HPLC) and the most recent radioimmunoassay (RIA) techniques. This review attempts to provide awareness of the limitations of these methods and establishes the base for eventual development of a B12 reference method in our laboratory.

Binding, Competitive↗