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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

Identification of glutathione conjugates and mercapturic acids of 1,2-dibromopropane in female BALB/c mice by liquid chromatography-electrospray ionization tandem mass spectrometry.

Based on recent results that 1,2-dibromopropane (1,2-DBP) causes hepatotoxicity and immunotoxicity in female BALB/c mice as well as a reduction of hepatic glutathione levels, the possible formation of glutathione conjugates and mercapturic acids of 1,2-DBP was investigated in vivo in the present studies. The following four metabolites were identified in the liver at 12 h after treatment with 1,2-DBP, by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS): M1, 2-hydroxypropylglutathione; M2, 2-oxopropylglutathione; M3, N-acetyl-S-(2-hydroxypropyl)-L-cysteine; and M4, N-acetyl-S-(2-oxopropyl)-L-cysteine. Ions of individual conjugates were observed at m/z 366, 364, 222 and 220, respectively. Characteristic product ions at m/z 237, 217, 204 and 202 for the identification of M1, M2, M3 and M4 were observed, respectively. In the sera isolated from the same animals, only mercapturic acids (M3 and M4) were observed by LC-ESI/MS. When female BALB/c mice were treated orally with 1,2-DBP at doses of 150, 300 and 600 mg kg(-1) once for 12 h, the production of glutathione conjugates and mercapturic acids in liver was apparently dose dependent, as were the concentrations of them in sera. When the production of metabolites from 1,2-DBP was investigated in liver following oral treatment with 600 mg kg(-1) 1,2-DBP for 6, 12, 24 and 48 h, metabolite concentrations were greatest at the first time point (6 h). The results explain the authors' previous studies that oral treatment with 1,2-DBP reduces the hepatic content of glutathione.

Acetylcysteine↗

Characterization of the Phase II metabolites of rutaecarpine in rat by liquid chromatography-electrospray ionization-tandem mass spectrometry.

From the authors' previous studies on the Phase I metabolism of rutaecarpine, nine metabolites formed were identified as products of hydroxylation on the aromatic rings in rat liver microsomes. In order to determine the possible metabolic fate of rutaecarpine, the Phase II metabolites of rutaecarpine were characterized in the present study by using liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS). When male Sprague-Dawley rats were treated intravenously with 4 mg kg(-1) rutaecarpine, 16 different Phase I and II metabolites were identified in urine including four sulfate and four glucuronide conjugates. Phase I metabolites of rutaecarpine were identified as four mono-hydroxylated metabolites (M2-5) and four isobaric di-hydroxylated metabolites (M6-9). These metabolites were identical to the in vitro metabolites except one, which was hydroxylated in the aliphatic moiety. In addition, Phase II metabolites were identified as conjugated with sulfate (S1-4) and glucuronide (G1-4). In faeces, 11 different metabolites were identified. The metabolites M8 and glucuronide conjugated (G1-4) were not detected. Structures of all metabolites were confirmed with CID fragmentation spectra of MS(2), MS(3) and retention times by LC/ESI-MS.

Alkaloids↗

Radon concentrations in groundwater in Busan measured with a liquid scintillation counter method.

In this paper, we present the first measurement of radon concentrations in drinking groundwater from private and public deep-bored wells located in the south-eastern area of Korea. The measurements were carried out on 439 samples by using a liquid scintillation method. The results show that the radon concentrations of the samples range from 0 to about 300 Bq l(-1). We find that Sasang ward shows the highest median value of radon concentration among 13 different wards, while Jung ward has the lowest. We find that the radon concentrations are highly dependent on the type of geological rock aquifers.

Environmental Monitoring↗

Polychlorinated biphenyl mixture aroclor 1254-induced oxidative stress plays a role in dopaminergic cell injury.

Oxidative stress (OS) is thought to participate in the pathogenesis of neurodegenerative disorders, including Parkinson's disease (PD). Excessive reactive oxygen species (ROS) production can occur during the normal aging process or following exposure to environmental toxicants. Dopamine neurons, which degenerate during PD, are particularly sensitive to oxidative stress. Polychlorinated biphenyls (PCBs), persistent and widespread pollutants, have been shown to adversely impact dopaminergic (DAergic) pathways, but the role ROS play in neurotoxicity remains unclear. To test the hypothesis that PCB exposure compromises dopamine neurons by stimulating ROS production, the direct toxicity and oxidative stress response following PCB exposure was examined both in MN9D dopamine cells and primary mesencephalic cultures. PCBs induced a time- and concentration-dependent increase in ROS production, which preceded cytotoxicity. Whereas intracellular GSH depletion exacerbated PCB effects, antioxidant pretreatment attenuated ROS production and cell death. Coincident alterations in antioxidant defense enzymes also accompanied ROS production, including decreased MnSOD and increased CuZnSOD protein levels. The robust elevation in heme oxygenase-1 levels further support the activation of oxidative stress mechanisms following PCB exposure. Furthermore, PCBs produced concentration-dependent reductions in intracellular dopamine levels and elevated dopamine turnover. Although the intracellular source of ROS remains unknown, these results suggest that sublethal PCB concentrations activate an oxidative stress-related pathway, which potentially disrupts dopamine neuron function.

Animals↗

Magnetic resonance imaging of metaplastic carcinoma of the breast: sonographic and pathologic correlation.

Metaplastic carcinoma of the breast is a rare disease. We describe the MRI findings with the correlative sonographic and pathologic features of two cases. On MRI, T2-weighted images demonstrate a relatively well-defined mass with high signal intensity cystic components. Dynamic enhancement subtraction images showed an early enhancing and delayed washout peripheral rim and non-enhancing internal components. A microlobulated, isoechogenic mass with cystic components was seen sonographically, and was histopathology related to necrosis and cystic degeneration. Although these features are not unique, metaplastic carcinoma should be included in the differential diagnosis for breast masses.

Adult↗

Somatic embryogenesis and plant regeneration from seeds of wild Dicentra spectabilis (L.) LEM.

Regeneration via somatic embryogenesis from callus was studied in Dicentra spectabilis. To obtain somatic embryogenic callus, we cultured D. spectabilis seeds on MS basal media supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D). The highest percentage of embryogenic callus formation was observed on media containing 1.0 mg/l 2,4-D under dark conditions. Somatic embryogenesis was studied by transferring the callus onto MS basal medium containing different concentrations (0.0, 0.1, 0.5, 1.0, 2.0 mg/l) of KIN (kinetin) and/or BAP. Somatic embryogenesis on MS basal media with 1.0 mg/l of KIN was excellent under light conditions. Somatic embryos were rooted by transferring them to half-strength MS basal media containing 2 g/l Phytagel. About 64.2% of the somatic embryos converted to rooted plantlets, 4% showed secondary embryogenesis and 31.8% did not develop and died. Rooted plantlets showed a 46% survival rate when acclimatized ex vitro.

2,4-Dichlorophenoxyacetic Acid↗

In vitro expression and pharmacology of the 5-HT7-like receptor present in the mosquito Aedes aegypti tracheolar cells and hindgut-associated nerves.

We have previously reported the cloning of a 5-hydroxytryptamine receptor (Aedes 5-HT7-like receptor) from adult Aedes aegypti. For functional expression of the Aedes 5-HT7-like receptor, CHO-K1 cells were stably transfected with a receptor expression construct, pC5-HT7. The Aedes 5-HT7-like receptor positively coupled to Gs protein, increasing intracellular cAMP in response to 5-HT; adenylyl cyclase activity was induced in a concentration-dependent, saturable manner. Only 5-HT, and not octopamine, dopamine or tyramine, caused the induction of cAMP. At 10 nM 5-HT a weak synergism was observed between octopamine and 5-HT. Other known agonists of the mammalian 5-HT7 receptor were tested. Their order of potency was: 5-HT >> 5-CT = 8-OH-DPAT >> pimozide. This is the first report on the functional expression of a mosquito neurohormone receptor.

Adenylyl Cyclases↗

Programs and procedures for a large x-ray safety program.

As radiation safety surveys of x-ray equipment are the responsibility of many health physicists, actual procedures vary among facilities. In an effort to share survey methodology of a large research institution, aspects of the x-ray safety program at Los Alamos National Laboratory are described. A definition of an intentional x-ray generating device is proposed in light of the absence or inadequacy of such a definition in applicable federal law and national consensus guidance documents. Regulations and procedures are discussed, and a sample database used in managing the Los Alamos program is presented.

Data Collection↗

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↗