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D Wang

Publications and source records attributed to D Wang.

At least 649 records · Page 36Linked to original sources
PubMed1998

The changes and significance of c-kit expression in the irradiated bone marrow of mice.

There have been only a few studies on the expression of c-kit in bone marrow after radiation and radiation-induced apoptosis of hematopoietic cells. LACA mice were examined after irradiation with a dose of 60Co gamma-rays. Using a light microscope, electron microscope, and immunohistochemistry, we studied radiation-induced apoptosis of hematopoietic cells and the expression of c-kit. We found that 5.5 Gy total body irradiation was a sublethal dose for LACA mice, induced suppression of leukocytes in blood, and of nucleated cells in bone marrow. Apoptosis of the hematopoietic cells occurred within 1 day after irradiation. The expression of c-kit was related to the recovery of hematopoiesis and played an important role in early hematopoiesis.

Animals↗
PubMed1998

Interleukin-3 gene expression in irradiated mouse bone marrow.

The expression of endogenous interleukin-3 (IL-3) gene in mouse bone marrow after whole body irradiation was assessed by immunocytochemistry, in situ hybridization (ISH), and in situ reverse transcription PCR (IS RT-PCR). The results showed obvious injury to the bone marrow followed by recovery 4 weeks after radiation. IL-3 protein was increased in the cytoplasm of recovering hematopoietic cells, especially at 21 days after radiation, while the mRNA was negative, except at 15 days when the mRNA was weakly positive by ISH. IS RT-PCR showed that IL-3 mRNA was strongly positive in the cytoplasm of recovering hematopoietic cells, especially at 10 to 15 days after radiation. We postulate that endogenous IL-3 gene may accelerate hematopoietic recovery in irradiated bone marrow and that IS RT-PCR may objectively report the regulation of IL-3 gene expression.

Animals↗
PubMed1998

Kinetic alterations of angiotensin-II and nitric oxide in radiation pulmonary fibrosis.

We studied the kinetic alterations of angiotensin-II (A-II) and nitric oxide (NO) in radiation pulmonary fibrosis (RPF) to determine the roles of these two types of vasoactive substances in the pathogenesis of RPF. We irradiated the right hemithorax of male Wistar rats with single doses of 0, 15, and 30 Gy of 60Co gamma rays and we examined the lung parenchyma at 1, 3, 5, and 7 months following the radiation. The rats were killed at the stated intervals and samples were obtained from the right lung. We measured types I and III procollagen mRNA by in situ hybridization and demonstrated the synthesis and distribution of A-II in the pulmonary tissue by immunohistochemistry. The formation and kinetic alterations of types I and III collagen were analyzed under polarized light microscope using Sirius Red stain. The hydroxyproline (Hyp) content was measured in the pulmonary tissue after digestion with HCl. A-II radiation immunoactivity (RIA) was assayed in pulmonary tissue homogenate. Pulmonary NO content, NO synthase (NOS), and the angiotensin converting enzyme (ACE) activities were also measured. Our results showed that types I and III collagen genes began to be expressed 1 month after irradiation. Type I collagen gene increased significantly, reaching its peak 3 months after irradiation. As the irradiation dosage was increased from 15 to 30 Gy, the type I collagen gene content increased significantly, while type III significantly decreased. The Hyp content increased with the passage of time after irradiation. Pulmonary A-II RIA increased significantly with the dose of irradiation and was chiefly produced by fibroblasts and macrophages in the interstitium, bronchiolar epithelium, and the anteriolar wall. Pulmonary NO and NOS activities decreased following irradiation. One month following irradiation, the expression of the type I collagen gene begins to increase, with a significant increase in both Hyp and type I collagen 3 months after irradiation. The histogenesis of RPF may be related to A-II. The interstitial cells, the bronchiolar epithelium, and the arteriolar wall can produce A-II and need not pass through the ACE pathway. Our results suggest that the A-II increase and NO decrease may have a role in the pathogenesis of RPF.

Angiotensin II↗
PubMed1998

The protective action of taurine and L-arginine in radiation pulmonary fibrosis.

Taurine has the ability to resist the action of oxygen-free radicals by protecting cytomembranes. L-arginine increases the nitric oxide in the organism, inhibiting some types of abnormal cell proliferation. We examined the actions and protective mechanisms of these two substances against radiation pulmonary fibrosis. The mRNA transcription and the distribution of pulmonary types I and III procollagen were measured by in situ hybridization. Type I and type III collagen in the interalveolar septa and the change ratio between the two types were assayed using Sirius Red staining and polarizing microscopy. Angiotensin converting enzyme (ACE), hydroxyproline (Hyp), malondialdehyde (MDA), and nitric oxide (NO) were measured in lung tissue after irradiation. We found that, following administration of taurine or arginine, the mRNA transcription of procollagen types I and III significantly decreased in the pulmonary tissue 3 months after irradiation and the Hyp content was lower, especially after the administration of arginine. The extent of the radiation-induced decrease in ACE activity was markedly attenuated. As a substrate of NO production, arginine can significantly increase the pulmonary NO content. We conclude that the administration of taurine or arginine can significantly decrease the mRNA transcription of procollagen and the synthesis of type I collagen in the interalveolar septa and can decrease the Hyp content of the pulmonary tissue. Following irradiation, NO can attenuate the radiation-induced decrease in ACE activity. Our study suggests that exogenous NO and taurine significantly protect against radiation pulmonary injury.

Animals↗
PubMed1998

Preventive effects of selenium-enriched spiruline (SESP) on radiation pneumonitis.

We studied the early effects of selenium-enriched spiruline (SESP) on radiation-induced pulmonary fibrosis using histopathology, cDNA-mRNA dot-blot hybridization, and a biochemical technique for quantifying the hydroxyproline content. We found marked lung injury in the irradiated group, whereas only slight hyperemia, hemorrhage, exudation, and thickness of the interalveolar wall in the lung in the selenium-treated group. The hydroxyproline (hyp) content in the lungs and the synthesis of type III collagen mRNA decreased in the SESP-treated group. Selenium-enriched spiruline may have a protective effect on the development of radiation pneumonitis.

Animals↗
PubMed1998

CT findings in splenic tuberculosis.

Splenic tuberculosis is rare and delay in diagnosis is common. We present two cases of splenic tuberculosis with their appearance on CT. The CT findings were multiple, round or ovoid, low-density lesions without calcification. Except for non-specific lymphadenopathy no abnormality nor history suggestive of tuberculosis was revealed prior to laparotomy. Although CT does not confirm a diagnosis of splenic tuberculosis, it would be a valuable examination to define the extent of disease and guide aspiration biopsy.

Abscess↗
PubMed1998

Selenium and arsenic in the environment in Finland.

A characteristic feature of glaciated Precambrian environments is their low selenium content, as a chalcophile element, Se, replaces sulfur in many of the sulfide minerals, for example, pyrite, chalcopyrite, pyrrhotite, and pentlandite. The average Se concentration in rocks and related till deposits in Finland is in the range of 0.01 to 0.2 mg/kg. Due to geological conditions, Se concentrations in surface and ground water are low in Finland compared with other countries. In a nationwide study dealing with the hydrogeochemistry of headwater streams, the median Se concentration in streams during August to September 1990 was 30 to 180 microg/L. For comparison, Se concentrations in shallow well waters are generally in the range of 50 to 1000 microg/L. The Se concentrations in stream sediments varied from 0.03 to 3.94 mg/kg. There was a highly significant correlation between the Se concentrations in stream water and in stream sediment. The streams with Se concentrations exceeding the general level in both water and sediment were most common in southern Finland. A speciation study on Finnish stream waters revealed that there were equal proportions of Se complexed with humic substances (36%) and Se as a selenate species (36%), whereas selenite accounted for less than 10% of total Se. About 8% of the Se in stream water occurred in particulate form. In an effort to enhance the Se intake of Finns through diet, Se-supplemented fertilizers have been used nationwide since 1985. While greatly improving Se levels in the population, the measure has raised concerns about undesirable environmental effects. Therefore, the amount of Se added to fertilizers has been reduced since 1991. Differing in behavior from Se, arsenic is considered one of the most toxic metals derived from the natural environment. Alarm has been triggered in Finland by the recent lowering from 50 microg/L to 10 microg/L of the upper level of As permissible in potable water, the recent information of high As concentrations in water from drilled bedrock wells, and the findings of international medical studies suggesting that As is a carcinogen. The most important source of As is arsenopyrite (FeAsS). Hence, high As concentrations most frequently occur in areas of sulfide mineralization, often in connection with occurrences of mafic rocks such as gabbros, amphibolites, and peridotites. The As concentrations in till fines, the most common glaciogenic soil type in Finland, reflect those in bedrock. The concentrations in groundwater are controlled by the chemical composition of the bedrock and the soil and prevailing hydrogeochemical conditions, for example, pH and Eh levels. Arsenic concentrations are lowest in surface water and swiftly flowing shallow ground water discharged by springs and are somewhat higher in shallow wells dug into overburden. By far, the highest As concentrations are to be found in wells drilled into bedrock (maximum 1 to 2 mg/L), although the concentrations vary by several orders of magnitude from well to well. The highest probability of encountering deleteriously arsenious well water is in areas with characteristic As anomalies in the till and bedrock. Hence, it is important to understand local geological conditions, particularly in the case of wells drilled into bedrock. The risk of deleteriously high As concentrations occurring in captured springs and shallow wells is slight.

Animals↗
PubMed1998

Family and sociodemographic influences on patterns of leaving home in postwar Britain.

We identify child-level and parent-level characteristics associated with children's patterns of leaving home. We use a multilevel discrete-time hazards model to examine the impact of family and demographic factors at both levels, and utilize the Alternating Conditional Expectation algorithm optimally to transform the dependent and independent variables. We find that measured variables at both the child and the parent level have important influences, as do period and cohort factors. However, unmeasured parent-level factors have an influence on the departure of children that is broadly similar in magnitude to measured factors.

Adolescent↗
PubMed1997

Chemosignal transduction in the vomeronasal organ of garter snakes: Ca(2+)-dependent regulation of adenylate cyclase.

Earthworm shock secretion contains a 20-kDa vomeronasally mediated chemoattractive protein for garter snakes. Both the ligand-receptor binding and the chemoattractivity of ES20 are Ca(2+)-dependent. When ES20 binds to its G-protein-coupled receptors in the vomeronasal epithelium, the inositol 1,4,5-trisphosphate (IP3) level is increased, but the level of cAMP is reduced. Furthermore, forskolin-stimulated levels of cAMP are completely blocked by ES20-receptor binding or by Ca2+ alone and the effect of calcium ions can be nullified by EGTA. Previously, we hypothesized that the decrease in cAMP was due to activation of a Ca(2+)-dependent phosphodiesterase. In the present study, we provide evidence that the decrease in cAMP is due mainly to the regulation of adenylate cyclase (AC) activity by Ca2+ or is indirectly mediated by ES20. Results obtained with intact vomeronasal sensory epithelium suggest that the binding of ES20 to its receptors facilitates generation of IP3 which mobilizes intracellularly sequestered Ca2+, resulting in an increase of cystosolic Ca2+. A further increase in cytosolic Ca2+ occurs through Ca2+ influx from extracellular sources. Garter snake vomeronasal AC does not require calmodulin for its activity and shows a biphasic response to increasing concentrations of Ca2+; its activity is modulated both positively and negatively by this bivalent cation.

Adenylate Cyclase Toxin↗
PubMed1997

Cloning and expression of a gene encoding a protein obtained from earthworm secretion that is a chemoattractant for garter snakes.

The protein ES20, derived from earthworm shock secretion, is a vomeronasally mediated chemoattractant for garter snakes (Jiang, X. C., Inouchi, J., Wang, D., and Halpern, M. (1990) J. Biol. Chem. 265, 8736-8744). Based on its 15-residue N-terminal amino acid sequence, degenerative oligodeoxynucleotide probes were synthesized and used to screen a cDNA library that was constructed in sense orientation using a Uni-ZAPTM XR vector and XL1-Blue MRF' host. A gene was cloned from a polymerase chain reaction as well as from the cDNA library. A combination of the forward degenerative primer and T7 primer was used to obtain gene-specific DNA fragments, from which probes were synthesized and successfully used in screening the cDNA library. The ES20 gene is about 700 base pairs long and encodes 208 amino residues. The ES20 gene was excised from a recombinant plasmid pSK-ES20, ligated to pQE30 expression vector, and transformed into Escherichia coli strain JM109. The selected recombinant plasmids were transformed into expression host cell, E. coli M15[pREP4]. Three transformants were selected, induced with isopropyl-1-thio-beta-D-galactopyranoside for fusion gene expression and an expressed 20-kDa fusion protein purified under denaturing conditions. This protein was refolded and gave a positive reaction against ES20-specific polyclonal antibodies. The fusion protein that had not been denatured remained as an aggregate and was an active chemoattractant for garter snakes.

Amino Acid Sequence↗
PubMed1997

Upstream stimulatory factor binding to the E-box at -65 is required for insulin regulation of the fatty acid synthase promoter.

Fatty acid synthase (FAS) plays a central role in de novo lipogenesis in mammals. We have shown that FAS transcription rate is induced dramatically when fasted animals are refed with a high carbohydrate diet or when streptozotocin-diabetic mice are given insulin. We also reported that FAS gene transcription was up-regulated by insulin through the proximal promoter region from -71 to -50 and that upstream stimulatory factors (USFs), including USF1 and USF2, interact with this region in vitro. In the present study, by using site-directed mutagenesis of the -71/-50 region and correlating functional assays of the mutated promoter with USF binding activities, we demonstrate that the -65/-60 E-box motif (5'-CATGTG-3') is functionally required for insulin regulation and that USFs are in vivo components of the insulin response complex. Mutation of the -65/-60 E-box sequence abolished insulin response in both transiently and stably transfected 3T3-L1 adipocytes in the -2. 1 kb promoter context, which contains all the necessary regulatory elements of the promoter based on our previous transgenic mice studies, and in the minimal -67 promoter context. Gel mobility shift assays demonstrated that USFs can no longer bind to the -71/-50 promoter region when the E-box is mutated. Cotransfection of USF1 and USF2 expression vectors with the FAS promoter-luciferase reporter constructs increased insulin-stimulated FAS promoter activity. Moreover, cotransfection of dominant negative USF1 and USF2 mutants lacking the DNA binding domain inhibited the insulin stimulation of the FAS promoter activity. On the other hand, site-directed mutagenesis of the -65/-60 E-box surrounding sequences within the overlapped tandem copies of sterol regulatory element-binding protein (SREBP) binding sites prevented SREBP from binding to -71/-50 promoter region in vitro but had no effect on insulin regulation of the FAS promoter in vivo. When rat liver nuclear extracts were used in gel mobility shift assays, only USF-containing protein-DNA complexes that can be supershifted by specific USF antibodies were observed. These results demonstrate that upstream stimulatory factor binding to the E-box at -65 is required for insulin regulation of the fatty acid synthase promoter.

3T3 Cells↗
PubMed1997

HMG-I(Y) phosphorylation status as a nuclear target regulated through insulin receptor substrate-1 and the I4R motif of the interleukin-4 receptor.

Interleukin (IL)-4 is a cytokine that regulates both the growth and differentiation of hematopoietic cells. Its ligand binding specificity and important signal transduction mechanisms are conferred by the IL-4 receptor alpha chain (IL-4Ralpha). The I4R is a tyrosine-containing motif within IL-4Ralpha that is critical for proliferative responses to IL-4. Although the I4R also contributes to gene regulation, nuclear targets directly regulated by this motif have not been described. It is shown here that the tyrosine at position 497 in the I4R is critical for regulation of the phosphorylation status of a set of nuclear proteins that includes HMG-I(Y), small non-histone chromosomal proteins involved in the control of gene expression in hematopoietic cell lines. Moreover, IL-4 is unable to induce HMG-I(Y) phosphorylation in insulin receptor substrate-1-deficient cells, and the inhibitor wortmannin completely blocks IL-4 regulation of HMG-I(Y) phosphorylation status but not activation of an IL-4 Stat protein. Taken together, these data indicate that HMG-I(Y) is a nuclear target whose phosphorylation status is regulated through the I4R motif via insulin receptor substrate proteins, independent of activation of the Stat pathway.

Androstadienes↗
PubMed1997

An intercalation inhibitor altering the target selectivity of DNA damaging agents: synthesis of site-specific aflatoxin B1 adducts in a p53 mutational hotspot.

Aflatoxin B1 (AFB1) is a potent human carcinogen implicated in the etiology of hepatocellular carcinoma. Upon metabolic activation to the reactive epoxide, AFB1 forms DNA adducts primarily at the N7 position of guanines. To elucidate more fully the molecular mechanism of AFB1-induced mutagenesis, an intercalation inhibitor was designed to probe the effects of intercalation by AFB1 epoxide on its reaction with DNA. DNA duplexes were prepared consisting of a target strand containing multiple potentially reactive guanines and a nontarget strand containing a cis-syn thymidine-benzofuran photoproduct. Because the covalently linked benzofuran moiety physically occupies an intercalation site, we reasoned that such a site would be rendered inaccessible to AFB1 epoxide. By strategic positioning of this intercalation inhibitor in the intercalation site 5' to a specific guanine, the adduct yield at that site was greatly diminished, indicating that intercalation by AFB1 epoxide contributes favorably to adduct formation. Using this approach it has been possible to simplify the production of site-specifically modified oligonucleotides containing AFB1 adducts in the sequence context of a p53 mutational hotspot. Moreover, we report herein isolation of site-specifically AFB1-modified oligonucleotides in sequences containing multiple guanines. Use of intercalation inhibitors will facilitate both investigation of the ability of other carcinogens to intercalate into DNA and the synthesis of specific carcinogen-DNA adducts.

Aflatoxin B1↗
PubMed1997

Preferential interaction of the his pause RNA hairpin with RNA polymerase beta subunit residues 904-950 correlates with strong transcriptional pausing.

RNA secondary structures (hairpins) that form as the nascent RNA emerges from RNA polymerase are important components of many signals that regulate transcription, including some pause sites, all rho-independent terminators, and some antiterminators. At the his leader pause site, a 5-bp-stem, 8-nt-loop pause RNA hairpin forms 11 nt from the RNA 3' end and stabilizes a transcription complex conformation slow to react with NTP substrate. This stabilization appears to depend at least in part on an interaction with RNA polymerase. We tested for RNA hairpin interaction with the paused polymerase by crosslinking 5-iodoUMP positioned specifically in the hairpin loop. In the paused conformation, strong and unusual crosslinking of the pause hairpin to beta904-950 replaced crosslinking to beta' and to other parts of beta that occurred in nonpaused complexes prior to hairpin formation. These changes in nascent RNA interactions may inhibit reactive alignment of the RNA 3' end in the paused complex and be related to events at rho-independent terminators.

Amino Acid Sequence↗
PubMed1997

Cytogenetic effect of chronic low-dose, low-dose-rate gamma-radiation in residents of irradiated buildings.

BACKGROUND: Many people in Taiwan have been living in buildings constructed with cobalt-60-contaminated steel rods. To study the biological effects of chronic low-dose ionising radiation on the residents of one such building, micronucleus formation in these individuals was compared with that in controls. METHODS: The 73 residents had 77 age-and-sex-matched controls: 31 had 31 close relatives as controls (group A controls); eight of the 31 had a second set of close relatives; and the other controls were 38 residents in neighbouring buildings. Two micronucleus assays were used-a cytochalasin B (CBMN) assay and another involving incubation with cytarabine (CBMNA). Assay results are given as "frequency", or the number of binucleate cells containing one micronucleus per 1000 randomly examined binucleate cells. FINDINGS: The CBMN and CBMNA mean (SD) frequencies for 31 exposed individuals (0.016 [0.009] and 0.025 [0.013] respectively) were greater than those for their group A controls (0.009 [0.004] and 0.016 [0.009], respectively) (p = 0.0006 and 0.0002, respectively). The mean CBMN and CBMNA frequencies for all the exposed individuals (0.017 [0.011] and 0.030 [0.014], respectively) were significantly greater than those for all controls (0.011 [0.008] and 0.019 [0.01]; p = 0.0001 for both comparisons). The ranges of the differences in CBMN or CBMNA frequencies between 31 exposed individuals and their group A controls were 0.003 to 0.020 and 0.001 to 0.032, respectively. After adjustment for age, sex, and cigarette smoking, the adjusted relative risks of micronucleus formation from radiation exposure in all 73 residents was 1.58 (95% CI 1.42-1.71; p = 0.0001) by the CBMN assay and 1.64 (1.53-1.77; p = 0.0001) by the CBMNA assay. INTERPRETATION: These findings suggest that chronic low-dose and low-dose-rate gamma-ray environmental exposure may induce cytogenetic damage in human beings.

Adolescent↗
PubMed1997

Evidence that intramolecular interactions are involved in masking the activation domain of transcriptional activator Leu3p.

The Leu3 protein of Saccharomyces cerevisiae regulates the expression of genes involved in branched chain amino acid biosynthesis and in ammonia assimilation. It is modulated by alpha-isopropylmalate, an intermediate in leucine biosynthesis. In the presence of alpha-isopropylmalate, Leu3p is a transcriptional activator. In the absence of the signal molecule, the activation domain is masked, and Leu3p acts as a repressor. The recent discovery that Leu3p retains its regulatory properties when expressed in mammalian cells (Guo, H., and Kohlhaw, G. B. (1996) FEBS Lett. 390, 191-195) suggests that masking and unmasking of the activation domain occur without the participation of auxiliary proteins. Here we present experimental support for this notion and address the mechanism of masking. We show that modulation of Leu3p is exceedingly sensitive to mutations in the activation domain. An activation domain double mutant (D872N/D874N; designated Leu3-dd) was constructed that has the characteristics of a permanently masked activator. Using separately expressed segments containing either the DNA binding domain-middle region or the activation domain of wild type Leu3p (or Leu3-dd) in a modified yeast two-hybrid system, we provide direct evidence for alpha-isopropylmalate-dependent interaction between these segments. Finally, we use the phenotype of Leu3-dd-containing cells (slow growth in the absence of added leucine) to select for suppressor mutations that map to the middle region of Leu3-dd. The properties of nine such suppressors further support the idea that masking is an intramolecular process and suggest a means for mapping the surface involved in masking.

Amino Acid Sequence↗