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Biomedical subjects

D W Lee

Publications and source records attributed to D W Lee.

At least 19 recordsLinked to original sources

cDNA cloning and sequence analysis of the lectin genes of the Korean mistletoe (Viscum album coloratum).

We previously isolated a lectin of the Korean mistletoe (Viscum album coloratum). The cDNA clones that encode the A- or the B-chain of the Korean mistletoe lectin were cloned by reverse transcriptase polymerase chain reaction (RT-PCR). The mRNAs that were extracted from the Korean mistletoe were amplified, ligated into the pGEM-T easy vector, and screened with a Korean mistletoe lectin-specific probe. The probe was prepared by PCR amplification of the Korean mistletoe DNA using a primer set designed on the basis of amino acid sequences of the Korean mistletoe lectin that we had purified and reported. Unlike a recent report, which states that the European mistletoe lectin gene has no isoforms, several different clones of the A- and B-chains of the Korean mistletoe lectin were cloned from the same primer set. Three clones of each were selected for sequencing. The sizes of the A-chains were 762, 762, and 768 bp, respectively. The B-chain sizes were 798, 789, and 789 bp, respectively. Each of the clones showed significant variation in the amino acids sequence, including the N-linked glycosylation sites of the lectin. The sequence analysis of each of the Korean lectin clones, in comparison with the European mistletoe lectin and the other type II ribosome binding proteins, is discussed in the text. In addition, Southern blot analysis of the Korean mistletoe genomic DNA, restricted by different enzymes and hybridized with the lectin DNA, showed multi-bands, supporting the existence of multicopy genes or a gene family. These data suggest that heterogeneity of the mistletoe lectin is not only introduced by post-translational modifications, but also by expression of isotypes of the lectin genes.

Amino Acid Sequence↗

[D-LEU-4]-OB3, a synthetic leptin agonist, improves hyperglycemic control in C57BL/6J ob/ob mice.

We have recently shown that the activity of a synthetic peptide corresponding to amino acid residues 116-130 of secreted mouse leptin is contained in a restricted sequence at the amino terminus of the peptide, between residues 116-122 (Ser-Cys-Ser-Leu-Pro-Gln-Thr, OB3). Substitution of the Leu residue at position 4 of OB3 with its D-isomer ([D-Leu-4]-OB3) enhanced the ability of OB3 (1 mg/day, ip, 7 days) to reduce body weight gain, food and water intake, and serum glucose in female C57BL/6J ob/ob mice. In the present study, we have utilized a pair-feeding approach to demonstrate that the antihyperglycemic action of [D-Leu-4]-OB3 is not solely due to its effects on caloric intake. One group of female C57BL/6J ob/ob mice (n=6) was fed ad libitum, and two additional groups (n=6 per group) were allowed 3.0 g food/mouse daily, an amount previously determined to satisfy [D-Leu-4]-OB3-treated mice. At the end of the 7-day test period, vehicle-injected mice fed ad libitum were approximately 10% heavier than their initial body weights, while pair-fed mice injected with vehicle and [D-Leu-4]-OB3-treated mice lost 5% of their initial body weights. After 1 day of treatment, blood glucose was reduced by 20% in pair-fed vehicle-injected mice, and by 40% in mice given [D-Leu-4]-OB3. Food restriction reduced blood glucose throughout the 7-day study, but not to levels seen in wild-type nonobese C57BL/6J mice of the same sex and age. After 2 days of treatment with [D-Leu-4]-OB3, however, blood glucose was reduced to levels comparable to those seen in wild-type nonobese mice. [D-Leu-4]-OB3 also lowered serum insulin levels by 53% when compared to mice fed ad libitum. Neither pair-feeding nor [D-Leu-4]-OB3 treatment had any apparent effect on thermogenesis. These results suggest that [D-Leu-4]-OB3 exerts its effects on serum glucose not only by suppressing caloric intake, but also through a separate effect on glucose metabolism which may involve increased tissue sensitivity to insulin.

Animals↗

Induction of apoptotic cell death by a p53-independent pathway in neuronal SK-N-MC cells after treatment with 2,2',5,5'-tetrachlorobiphenyl.

Apoptotic cell death is an active process, which is a critical feature of the regulated development of multicellular organisms. Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants, some of which may be neurotoxic. This study investigates the 2,2', 5,5'-tetrachlorobiphenyl (PCB 52) induced apoptosis in human neuronal SK-N-MC cells, and the role of p53 in this response. Upon treatments with PCB 52, time- and concentration-dependent inhibition of the cell viability was observed. PCB 52 also caused apoptosis, as measured by cell morphology and DNA fragmentation. The capability of PCB 52 to induce apoptosis was associated with the proteolytic cleavage of specific target proteins, such as poly(ADP-ribose) polymerase (PARP) and beta-catenin proteins, suggesting the possible involvement of caspases. In general, DNA-damaging agents induce accumulation of the tumor suppressor protein p53, leading cells to either growth arrest in G1, or apoptosis. However, our data showed that both p53 and Bcl-2 protein levels were decreased in a time-dependent manner during apoptosis after exposure to PCB 52. These results suggest that PCB 52 induced a p53-independent apoptosis in these cells.

Apoptosis↗

Hippocampal volume does not change seasonally in a non food-storing songbird.

Seasonal differences in hippocampal morphology have been reported in food-storing birds. Non food-storing species have not been investigated however. It is therefore unclear whether seasonal changes in the hippocampus are specifically related to food-storing or reflect a more general seasonal mechanism that occurs in both food-storing and non food-storing birds alike. We determined the volumes of the hippocampal formation and remaining telencephalon in the non-storing male song sparrow (Melospiza melodies morphna) in two experiments comparing birds collected in the spring and fall of 1992-94 (Experiment 1) and 1997 (Experiment 2). Although pronounced seasonal changes in song control nuclei such as the HVC and RA were previously reported for the same brains used in Experiment 1, we found that hippocampal volume did not change with season in either Experiment 1 or 2 for these song sparrow brains. These results suggest that seasonal changes in the hippocampus do not occur in this non food-storing species and may be specific to food-storing birds.

Animals↗

Early clinical outcomes after subfascial endoscopic perforator surgery (SEPS) and saphenous vein surgery in chronic venous insufficiency.

BACKGROUND: Subfascial endoscopic perforator surgery (SEPS) has recently become popular as a minimally invasive way to treat chronic venous insufficiency (CVI) of the lower extremities. We report the early clinical outcomes of SEPS and saphenous vein surgery in a prospective series of Chinese patients who presented with severe CVI. METHODS: All patients referred to our hospital for the management of severe CVI (class IV disease or above) after January 1998 underwent SEPS using an ultrasonic scalpel in conjunction with saphenous vein surgery. All patients were followed up prospectively to assess ulcer healing, ulcer recurrence, and symptoms after SEPS. Clinical outcome was evaluated by the scoring system suggested by the Consensus Committee of the American Venous Forum on Chronic Venous Disease. RESULTS: Over a 24-month period, we performed 36 SEPS on 31 patients. Nineteen lower extremities (53%) had active or healing ulcers. Sapheno-femoral ligation was also performed in 33 limbs (92%). Four limbs (11%) developed superficial wound infection, and two (6%) had saphenous nerve dysesthesia. The mean clinical score and disability score decreased from 8.42 to 3.42 and 1.45 to 0.31 respectively, after a median follow-up of 14 months (range, 6-22) (p < 0.005). Eleven ulcers (58%) healed within 6 weeks after surgery. At 1-year follow-up, ulcer recurrence was found in two legs (11%). CONCLUSION: SEPS is safe and feasible. Early clinical results have shown a promising outcome in patients with severe chronic venous insufficiency.

Adult↗

Size analysis of automobile soot particles using field-flow fractionation.

Soot particles emitted from various automobile engines are analyzed for size distributions using field-flow fractionation (FFF). Soot samples are prepared for FFF analysis using a three-step procedure, where a layer of soot particles is focused between the layers of n-hexane and water, followed by dispersing of particles in water containing 0.05% Triton X-100. The mean diameters determined by FFF show similar trends with those obtained from dynamic light scattering (DLS) and scanning electron microscopy (SEM). Data from FFF are also compared with those from an on-line scanning mobility particle sizer (SMPS). SMPS size distributions extend further to larger size than those of FFF distributions, which indicates the three-step sample preparation procedure effectively disaggregates the agglomerated particles. Although the amount of particulate matter (PM) emitted from a heavy-duty diesel engine is much higher than that from a light-duty diesel engine, the size distributions of soot particles show no significant difference between heavy- and light-duty diesel engines. The engine-operating mode (engine speed and load rate) does not seem to affect significantly the size distribution of soot particles. It was found that the PM from a turbocharged diesel engine contains a higher percentage of particles smaller than 100 nm than an engine with a naturally aspirated (NA) air-inhalation system. As for gasoline engines, the PM collected after the catalytic converter has a narrower size distribution than those collected before and has a higher percentage of particles smaller than 100 nm.

Air Pollution↗

Effect of dominant follicle removal before superstimulation on follicular growth, ovulation and embryo production in Holstein cows.

This study was to investigate whether removing the dominant follicle 48 h before superstimulation influences follicular growth, ovulation and embryo production in Holstein cows. After synchronization, ovaries were scanned to assess the presence of a dominant follicle by ultrasonography with a real-time linear scanning ultrasound system on Days 4, 6 and 8 of the estrus cycle (Day 0 = day of estrus). Twenty-six Holstein cows with a dominant follicle were divided into 2 groups in which the dominant follicle was either removed (DFR group, n=13) by ultrasound-guided follicular aspiration or left intact (control group, n=13) on Day 8 of the estrus cycle. Superovulation treatment was initiated on Day 10. All donors were superovulated with injections of porcine FSH (Folltropin) twice daily with constant doses (total: 400 mg) over 4 d. On the 6th and 7th injections of Folltropin, 30 mg and 15 mg of PGF2alpha (Lutalyse) were given. Donors were inseminated twice at 12 h and 24 h after the onset of estrus. Embryos were recovered on Day 6 or 7 after AI. During superstimulation, the number of follicles 2 to 5 mm (small), 6 to 9 mm (medium) and > or = 10 mm (large) was determined by ultrasonography on a daily basis. At embryo recovery, the number of corpora lutea (CL) was also determined by ultrasonography and blood samples were collected for analysis of progesterone concentration. Follicular growth during superstimulation was earlier in the DFR group than in the control group. The number of medium and large follicles was greater (P < 0.01) in the DFR group than in the control group on Days 1 to 2 and Days 3 to 4 of superstimulation, respectively. The numbers of CL (9.6+/-1.1 vs 6.1+/-0.9) and progesterone concentration (30.9+/-5.4 vs 18.6+/-3.5 ng/mL) were greater (P < 0.05) in the DFR group than in the control group, respectively. The numbers of total ova (7.7+/-1.3 vs 3.9+/-1.0) and transferable embryos (4.6+/-0.9 vs 2.3+/-0.8) were also greater (P < 0.05) in the DFR group than in the control group, respectively. It is concluded that the removal of the dominant follicle 48 h before superstimulation promoted follicular growth, and increased ovulation and embryo production in Holstein cows.

Animals↗

On-line particle concentrator with upstream ultrafiltration in continuous SPLITT fractionation.

An upstream ultrafiltration (UU) method is employed for the on-line concentration of collected particle fraction and for the convenient regulation of flow rates in split-flow thin (SPLITT) fractionation. Concentration of the collected particle fraction is necessary to fractionate particles by smaller cutoff diameters in SPLITT fractionation. By introducing a simple device utilizing upstream ultrafiltration with tangential flow, particle solution collected at each SP

Journal Article↗

Specificity of lectin activity of Bacillus thuringiensis parasporal inclusion proteins.

Sheep erythrocyte-agglutinating parasporal inclusion proteins from four Bacillus thuringiensis strains (FITC-20, FITC-73, FITC-76 and IBC-1456) were examined for lectin activity against erythrocytes from four mammalian (rabbit, horse, cow and guinea pig) and an avian (chicken) species. Of the five erythrocyte species, only rabbit cells were agglutinated with the protein of the human cancer cell-killing strain IBC-1456. No haemagglutination (HA) activities were shown in other protein-erythrocyte combinations. The lectin activity of the strain IBC-1456 against rabbit cells was strongly inhibited by preincubation with D-galactose. Overall results revealed that the B. thuringiensis lectins have a preference for sheep erythrocytes.

Animals↗

Direct and precise determination of environmental radionuclides in solid materials using a modified Marinelli beaker and a HPGe detector.

A simple but precise detection method was studied for the determination of natural radionuclides using a conventional HPGe detector. A new aluminium beaker instead of a plastic Marinelli beaker was constructed and examined to reach radioactive equilibrium conditions between radon and its daughter elements without the escape of gaseous radon. Using this beaker fifteen natural radionuclides from three natural decay series could be determined by direct gamma-ray measurement and sixteen radionuclides could be determined indirectly after radioactive equilibrium had been reached. Analytical results from ground water were compared with those from conventional alpha spectroscopy and the results agreed well within 12% difference. Nitrogen gas purge was used to replace the surrounding air of the detector to obtain a stable background and reducing the interference of radon daughter nuclides in the atmosphere. The use of nitrogen purging and the aluminium Marinelli beaker results in an approximately tenfold increase of sensitivity and a decrease of the detection limit of 226Ra to about 0.74 Bq kg(-1) in soil samples.

Journal Article↗

Bacillus thuringiensis: a common member of microflora in activated sludges of a sewage treatment plant.

Bacillus thuringiensis was recovered at a high frequency from activated-sludge system environments in an urban sewage-digestive plant. All of the test materials, sampled at several digesting steps, contained the organism. Of 515 colonies belonging to the B. cereus/B. thuringiensis group, 45 (8.7%) were assigned to B. thuringiensis. The highest density of this bacterium was 1.6 x 103 cfu/ml in a scum sample of the first aeration basin. Among the 45 isolates, 7 were assigned to the known H serovars. Two isolates of the serovar kenyae isolates exhibited Lepidoptera-specific toxicity. Diptera-specific toxicity was shown by an isolate of serovar israelensis and a serologically undefined isolate. Lectin activity was associated with 12 isolates.

Animals↗

The IsdA gene is necessary for sexual development inhibition by a salt in Aspergillus nidulans.

Using one of 17 expressed sequence tags (ESTs) previously identified as specific to the late sexual developmental (LSD) stage of Aspergillus nidulans, a gene for the subject EST was isolated. The determined DNA sequences revealed an open reading frame encoding a 356 amino acid polypeptide which does not share a sequence similarity to previously identified genes or proteins. The isolated gene was named lsdA (late sexual development), since it was expressed abundantly at the LSD stage. The lsdA gene expression began earlier than at the LSD stage. Disruption of the lsdA gene in the veA+ background strongly induced sexual development under conditions where sexual development in wild-type strains is almost completely inhibited. In contrast, in the veA1 background, an lsdA null mutant failed to show any phenotypic difference in sexual development under the various conditions tested. These results indicate that the lsdA gene may be responsible for inhibiting the sexual development of veA+ strains by a high concentration of a salt.

Amino Acid Sequence↗

A study on the distribution of chlorination by-products (CBPs) in treated water in Korea.

Fifteen chlorination by-products were analyzed in 416 water samples collected from 35 water treatment plants in Korea from 1996 to 1998. These samples were divided into five groups according to water sources (Han-river, Nakdong-river, Youngsan-river, Kum-river and Cheju) and detected CBPs were classified into six classes (trihalomethanes; THMs, haloacetic acids; HAAs, haloacetonitriles: HANs haloketones; HKs, chloralhydrate; CH, chloropicrin; CP) and then, it was observed the detection tendency and frequency of CBPs in each water source. The total concentration of CBPs in treated water from Nakdong-river or Han-river was higher than those from the other rivers. And the distribution pattern of each class of CBPs was similar in all water sources. THMs were the highest portion in the range of 40-50%, and HAAs and HANs were 28-35 and 9-15%, respectively. And there was a strong correlation between HANs and HKs (r=0.813). Each and total concentrations of CBPs showed to be more affected by the water source in two-way analysis of variance (two-way ANOVA) among the concentration of CBPs, the source of water and season.

Acetates↗

Effect of estrogen on endothelial dysfunction in postmenopausal women with diabetes.

BACKGROUND: Treatment with estrogen causes increased release of nitric oxide and this appears to be an important mechanism involved in the cardioprotective effect of estrogen. Previous studies have clearly shown that estrogen has an acute vasodilatory effect. Recently, abnormality of flow mediated endothelial dilation, which is defined as an endothelial dysfunction, has been proposed as an early manifestation of atherogenesis. However, its effect in type 2 diabetes is unknown. Stimulus-provoked endothelium dependent dilatation, using high-resolution external vascular ultrasound, is a useful method in the measurement of endothelial function. The main advantages of this method are that it is completely non-invasive, accurate and reproducible. This classical method for the measurement of endothelial dysfunction, utilizing non-invasive, high-resolution ultrasonography, has some limitations especially in terms of reproducibility. To improve the accuracy and reproducibility of this method, measurements made for this study involved the use of an additional attachment. This device was an ultrasound probe that was attached to the arm of patient. In addition, this study evaluated not only a change in diameter in terms of flow mediated vasodilation, but also initial and peak response times during this test. The aims of this study are: (1) to evaluate the flow mediated peak vasoreactivity (FMD), the lag time from base line to initiation of vasodilation (IRT) and the peak response of endothelial dependent vasodilation in a control group and in young diabetic patients; and (2) to evaluate the effect of estrogen treatment on flow mediated vasodilation in postmenopausal women with diabetes. METHOD: Measurements were taken for flow mediated vasodilation (endothelial dependent vasodilation: FMD), endothelial independent vasodilation (EID) and lag time from base line to initial reaction of FMD. The aforementioned non-invasive method and attachment were used. Subjects for the experiment were 12 young people with diabetes (six male and six female, mean age 26.3) and 12 age-matched healthy controls (six male and six female, mean age 25.6). In addition, measurements were taken for FMD, EID and lag time in the brachial artery before and after estrogen supplementation (Premarin 0.625 mg for 7 days). Subjects for the experiment were 16 normal postmenopausal women and 18 age-matched postmenopausal women with type 2 diabetes. RESULT: There is no significant difference in BMI (body mass index), mean age, or blood pressure between the control group and the young diabetic group. There were no differences in age, total and LDL cholesterol levels or body mass index between the groups of postmenopausal women (P<0.05). However, the HDL cholesterol level was found to be significantly lower in postmenopausal women with diabetes than in normal postmenopausal women (respectively, a mean+/-SD; 38.95+/-11.96 mg/dl vs. 52.20+/-9.55 mg/dl, P<0.05). The FMD of young diabetic patients is slightly lower than that of age-matched young healthy controls (healthy control: 8.9+/-2.7%, young diabetic patients: 6.3+/-2.1%, P<0.05). There was no difference in endothelial independent vasodilation between the control group and the young diabetic group. The IRT of FMD was significantly shorter in the healthy control group than in the young diabetic group (healthy control: 20.4+/-2.8 s, diabetic group: 29.5+/-5.4 s, P<0.0001). In contrast, the IRT of EID was no different between the two groups. The IRT of FMD showed a significant negative correlation with FMD (r=-0.61, P<0.01) and a significant negative correlation with high-density lipoprotein (HDL) cholesterol (r=-0.68, P<0.0001). The basal endothelium dependent vascular reactivity was significantly decreased in postmenopausal women with diabetes compared with normal postmenopausal women (8.0+/-3.9 vs. 13.7+/-6.2%, P<0.05). Estrogen supplementation increased endothelium dependent vasodilation not only in postmenopausal women with diabetes (from 8.0+/-3.9 to 15.1+/-4.0%, P<0.05) but also in normal postmenopausal women (from 13.7+/-6.2 to 20.1+/-4.7%, P<0.05). In contrast, the responses to sublingual nitroglycerin were comparable between postmenopausal women with diabetes (from 21.1+/-6.0 to 22.1+/-4.1%, P>0.05) and normal postmenopausal women (from 25.8+/-7.8 to 25.2+/-4.5%, P>0.05), both before and after estrogen supplementation. CONCLUSION: These findings indicate that in addition to the percentage change in FMD, lag time from base line to initial response time (IRT) of FMD is a good indicator for evaluation of endothelial dysfunction. Also, this new parameter may be a more sensitive parameter for differentiation between the diseased state and the normal state of the endothelium than percentage changes in FMD. In terms of postmenopausal women, endothelial dysfunction was prominent in women with diabetes and was significantly improved by estrogen but not reversed. These results suggest that other factors in addition to estrogen deficiency play a role in endothelial dysfunction in postmenopausal women with diabetes mellitus.

Adult↗

Sclerotherapy with picibanil (OK-432) for congenital lymphatic malformation in the head and neck.

HYPOTHESIS/OBJECTIVES: Congenital lymphatic malformations of the head and neck (LMHN) present special challenges to the otolaryngologist-head and neck surgeon. Recently, a number of sclerotherapy trials have shown promising results. In this study, we present our experiences with picibanil (OK-432) sclerotherapy for this lesion. STUDY DESIGN: Retrospectively review. METHODS: We retrospectively reviewed 21 patients who have undergone sclerotherapy with picibanil for LMHN. RESULTS: Satisfactory response with complete or nearly complete shrinkage of the lesions was observed in 15 cases after repeated sclerotherapy (average, two times). We did not observe any significant morbidity or complications in the patients treated with picibanil. Reduction in size of the mass was achieved in weeks to months. Some of the patients who had not had any other previous treatment showed remarkable reductions in size even after the first therapy. When we used picibanil sclerotherapy as a primary treatment for the LMHN, most of our patients showed satisfactory results regardless of the size or location of the lesions. CONCLUSION: Given with our experience and the reports that failure of picibanil sclerotherapy does not hinder subsequent surgical salvage procedures, we recommend trying picibanil sclerotherapy as a primary treatment for the LMHN and performing surgical excision as a secondary modality if the response to the sclerotherapy is not satisfactory.

Child↗

Thermoanaerobacter yonseiensis sp. nov., a novel extremely thermophilic, xylose-utilizing bacterium that grows at up to 85 degrees C.

A novel strictly anaerobic, extremely thermophilic, spore-forming and xylose-utilizing bacterium, designated strain KB-1TP (type and patent strain), was isolated from a geothermal hot stream at Sileri on Java island, Indonesia. The cells were rod-shaped, motile and had terminal spores. The newly isolated strain stained gram-positive and the cells occurred singly or in pairs during the exponential growth phase. The temperature optimum for growth was 75 degrees C and growth occurred in the range 50-85 degrees C. The pH range for growth was 4.5-9.0, with an optimum at pH 6.5. Strain KB-1TP grew chemo-organotrophically by fermenting a wide range of substrates such as glucose, fructose, D-xylose, lactose, maltose, sucrose, mannose, galactose, cellobiose, pullulan and soluble starch. Arabinose, xylan, cellulose, olive oil and Tween 80 were not fermented. The predominant fermentation end products after growth on glucose were lactate, acetate, ethanol, CO2 and small amounts of isovaleric acid, butyric acid, propionic acid, 1-pentanol and 2-propanol. Thiosulfate was reduced to H2S. Strain KB-1TP was sensitive to tetracycline, chloramphenicol, penicillin G, neomycin, kanamycin, vancomycin and rifampicin at concentrations of 100 microg ml(-1). No effect was observed with chloramphenicol and neomycin at concentrations of 10 microg ml(-1). This indicates that strain KB-1TP belongs to the bacterial domain. The G+C content of the DNA was 37 mol%. The comparison of the 165 rDNA sequence to that of closely related strains revealed that strain KB-1TP belonged to clostridial cluster V, showing highest sequence identities (92.7%) to members of the genus Thermoanaerobacter. Taking into account the physiological and molecular properties of the new isolate, it is proposed that strain KB-1TP should be classified as a new species of the genus Thermoanaerobacter, designated Thermoanaerobacter yonseiensis. The type strain, KB-1TP, has been deposited in the Korean Federation of Culture Collections (KFCC 11116P) as a patent strain and in the Deutsche Sammlung von Mikroorganismen und Zellkulturen as a type strain (= DSM 13777T).

Bacillaceae↗