Search PubMed⌕ Search

Biomedical subjects

D Wang

Publications and source records attributed to D Wang.

At least 127 records · Page 7Linked to original sources
PubMed2001

Protein tyrosine kinase Csk-catalyzed phosphorylation of Src containing unnatural tyrosine analogues.

Using expressed protein ligation, five unnatural tyrosine analogues (amino-phenylalanine, homotyrosine, 2-methyl-tyrosine, (alphaS,betaR)-beta-methyl-tyrosine, and 2,6-difluoro-tyrosine) were incorporated into Src in place of the natural tail tyrosine residue. These semisynthetic substrates were evaluated as Csk substrates or allosteric activators. It appears that the tyrosine phenol hydroxyl is unlikely to be contributing significantly to Src's ground-state binding affinity for Csk. It has been observed that stabilizing tyrosine conformers can further optimize Src's already high substrate efficiency. These latter findings contrast similar studies with synthetic peptide substrates and highlight the value of investigation of protein kinase substrate selectivity with protein substrates.

CSK Tyrosine-Protein Kinase↗
PubMed2001

Fractal and chaotic behavior of circular cellular automata.

A new type of circular cellular automata (CCA) has been introduced. The evolutions of the CCA obtained by the clockwise, anticlockwise, and scanning line-by-line site sequence in the successively growing rings divided from a square lattice are studied. The evolution seems to form a twisty fishnet when the CCA are grown by the first two sequences. Sierpinski triangle gasket or the modulated ones are formed in the fourth quadrant of the CCA grown by the line scanning sequence. Fractal analysis is used to characterize the relationships between the pattern formed and the initial position of the growing ring and it is found that the pattern is very sensitive to the initial growth condition, showing the chaotic behavior.

Journal Article↗
PubMed2001

Synthetic utility of stannyl enolates as radical alkylating agents.

[reaction: see text] The radical-initiated beta-ketoalkylation of haloalkanes with tributylstannyl enolates is described. Stannyl enolates derived from aromatic ketones are reactive toward the homolytic beta-ketoalkylation of simple haloalkanes as well as those activated by an electron-withdrawing group. The reactivity of stannyl enolates as radical alkylating agents can be utilized for an efficient three-component coupling reaction among stannyl enolates, haloalkanes, and electron-deficient alkenes.

Journal Article↗
PubMed2001

Lack of association between the tryptophan hydroxylase gene A218C polymorphism and attention-deficit hyperactivity disorder in Chinese Han population.

Previous studies have suggested that the serotonergic (5-HT) system might be involved in the development of Attention-deficit hyperactivity disorder (ADHD). ADHD is frequently characterized by aggressive and impulsive behavior, a major symptom associated with reduction in serotonergic function. The tryptophan hydroxylase (TPH) gene is a reasonable candidate for ADHD because it encodes the rate-limiting enzyme in the process of 5-HT biosynthesis. In this study, we examined the relationship between the A218C polymorphism in TPH gene and ADHD. Sixty-nine ADHD patients and their biological parents were investigated. The A218C polymorphism in intron 7 of TPH gene was detected by PCR-RFLP method. No allele or genotype concerned with this A218C polymorphism was found to be associated with ADHD when analyzed with the haplotype relative risk method. Therefore, our data indicate that the TPH gene A218C polymorphism may not be a susceptibility factor of ADHD in the Chinese Han population.

Alleles↗
PubMed2001

Protein kinase C epsilon suppresses Abeta production and promotes activation of alpha-secretase.

Deposition of plaques containing Abeta is considered important in the pathogenesis of Alzheimer's disease. Phorbol esters that activate protein kinase C (PKC) promote alpha-secretase-mediated processing of the beta amyloid precursor protein (APP), which generally reduces formation of Abeta. To determine which PKC isozymes mediate this process, we studied CHO cells that express human APP751. Phorbol 12-myristate, 13-acetate (PMA)-stimulated APP secretion, which was reduced by a general PKC inhibitor bisindoylmaleimide I, but not by Gö 6976, which inhibits PKCalpha, beta, gamma, and mu. Since PKCdelta and epsilon were the only other PMA-sensitive isozymes present, we studied cells that express selective peptide inhibitors of these isozymes. Expression of the PKCepsilon inhibitor inhibited PMA-induced APPs secretion and suppression of Abeta production. In contrast, the PKCdelta inhibitor had no effect. These results provide evidence that PKCepsilon decreases Abeta production by promoting alpha-secretase mediated cleavage of APP.

Alzheimer Disease↗
PubMed2001

[The effect of focal mild hypothermia on the temperature gradient in brain].

OBJECTIVE: Through studying the effect of focal mild hypothermia on the distribution of brain temperature, which reveals the function that the semi-conductor refrigerator apparatus decreases the temperature to mild hypothermia level during therapeutic time window. METHODS: Directly measure the intracerebral temperatures of different regions when we refrigerator the brains of piglets. According to the results of the measure and the formula of heat conduction, we can establish the three-dimensional space and the temperature gradient mathematical model of pig brain. RESULTS: (1) The theoretical data deduced by function model fit very well to the laboratory data, absolutely average error is 0.15 degree C. (2) After 17 minutes of the piglet refrigerated at 2-10 degrees C, thalamic and putamen temperature decreases to 32-34 degrees C. CONCLUSION: Apply the regional semi-conductor refrigerator apparatus to decrease the brain temperature to mild hypothermia level during therapeutic time window and find the function relationship between refrigerator temperature and brain temperatures so as to provide the theoretical proof for clinical application of focal mild hypothermia.

Animals↗
PubMed2001

[Risk factors of lung cancer and relevant comprehensive preventive strategy--a cohort study in Xuanwei, Yunnan Province, China].

OBJECTIVE: To systematically analyze the risk factors of lung cancer in Xuanwei, Yunnan Province, an area with a high prevalence of lung cancer, and in view of the above to propose a comprehensive preventive strategy. METHODS: A cohort study of 42,434 Xuanwei farmers who were born 1917-195, and still alive on 1st January 1976 was carried out from 1992 to 1996 with a follow-up time of 21 years. The data collection was done in two steps. A retrospective cohort study was conducted in the form of questionnaire survey in 1992 to understand the health situation of the interviewees from 1st January 1976 to 31st December 1991. In the second stage, a prospective cohort study was conducted to review the death records of the interviewees from 1st January 1992 to 31st December 1996. The data thus collected were analyzed by the Mantel-Haenszel analysis method, COX model and risk state analysis method. RESULTS: Indoor air pollution caused by biturminous coal burning is the major risk factor of lung cancer in Xuanwei. Stove improvement decreased the lung cancer mortality rate in both male and female subjects, but its beneficial can be recognized at least 10 years after. Smoking is another important risk factor of lung cancer in males in Xuanwei. In course of a long time since stove improvement smoking will become the major risk factor of lung cancer. The risky factors can be arranged in sequence of riskiness from high to low as bituminous coal burning, family history of lung cancer, personal history of chronic bronchitis, and smoking in males; and bituminous coal burning, family history of lung cancer, and personal history of chronic bronchitis in females. The risk state classification model established upon the basis of risk state analysis can be used in identifying effectively high risk population quantitatively for local administrators of health as well as for local residents. CONCLUSION: According to the comprehensive strategy for lung cancer control in Xuanwei, priority should be given to primary prevention with three steps: firstly to further reduce indoor air pollution by means of stove improvement and ventilation improvement, secondly strengthen smoking cessation, and thirdly to carry out chemoprevention and susceptibility study of lung cancer among high risk populations.

Air Pollution, Indoor↗
PubMed2001

Identification and characterization of a novel extracellular matrix protein nephronectin that is associated with integrin alpha8beta1 in the embryonic kidney.

The epithelial-mesenchymal interactions required for kidney organogenesis are disrupted in mice lacking the integrin alpha8beta1. None of this integrin's known ligands, however, appears to account for this phenotype. To identify a more relevant ligand, a soluble integrin alpha8beta1 heterodimer fused to alkaline phosphatase (AP) has been used to probe blots and cDNA libraries. In newborn mouse kidney extracts, alpha8beta1-AP detects a novel ligand of 70-90 kD. This protein, named nephronectin, is an extracellular matrix protein with five EGF-like repeats, a mucin region containing a RGD sequence, and a COOH-terminal MAM domain. Integrin alpha8beta1 and several additional RGD-binding integrins bind nephronectin. Nephronectin mRNA is expressed in the ureteric bud epithelium, whereas alpha8beta1 is expressed in the metanephric mesenchyme. Nephronectin is localized in the extracellular matrix in the same distribution as the ligand detected by alpha8beta1-AP and forms a complex with alpha8beta1 in vivo. Thus, these results strongly suggest that nephronectin is a relevant ligand mediating alpha8beta1 function in the kidney. Nephronectin is expressed at numerous sites outside the kidney, so it may also have wider roles in development. The approaches used here should be generally useful for characterizing the interactions of novel extracellular matrix proteins identified through genomic sequencing projects.

Alkaline Phosphatase↗
PubMed2001

Characterization of volatile organic compounds in smoke at municipal structural fires.

The objective of this study was to characterize volatile organic compounds (VOCs) found at municipal structural fires in order to identify sources of long-term health risks to firefighters, which may be contributing factors in heart disease and cancer. Firefighters collected air into evacuated Summa canisters inside burning buildings at nine municipal structural fires under conditions where they judged that at least some firefighters might remove their self-contained breathing apparatus masks. Volatile organic compounds were identified and quantified for 144 target compounds using cryogenic preconcentration and gas chromatography/mass spectral detection (GC/MSD) methodology operating in selected ion monitoring mode. Samples were also analyzed in SCAN mode and examined for the appearance of substances that were not present in the instrument standard calibration mixture. The spectra of municipal structural fires were surprisingly similar and remarkable for their simplicity, which was largely due to the dominating presence of benzene along with toluene and naphthalene. Propene and 1,3-butadiene were found in all of the fires, and styrene and other alkyl-substituted benzene compounds were frequently identified. Similar "fingerprints" of the same 14 substances (propene, benzene, xylenes, 1-butene/2-methylpropene, toluene, propane, 1,2-butadiene, 2-methylbutane, ethylbenzene, naphthalene, styrene, cyclopentene, 1-methylcyclopentene, isopropylbenzene) previously identified at experimental fires burning various solid combustible materials were also found at municipal structural fires, accounting for 76.8% of the total VOCs measured. Statistically significant positive correlations were found between increasing levels of benzene and levels of propene, the xylenes, toluene, 1-butene/2-methylpropene, 1,3-butadiene, and naphthalene. Given the toxicity/carcinogenicity of those VOCs that were found in the highest concentrations, particularly benzene, 1,3-butadiene, and styrene, further investigation of VOC exposures of firefighters is warranted. Benzene, or its metabolic product s-phenylmercapturic acid in urine, was identified as a suitable chemical marker for firefighter exposure to combustion products.

Alkenes↗
PubMed2001

First experimental observation on different ionic states of the tert-butoxy [(CH3)3CO*] radical.

A continuous tert-butoxy (CH3)+CO* radical beam is produced in situ by respective pyrolysis of both (CH3)3CONO at 115(+/-0.5) degrees C and (CH3)3COOC(CH3)3 at 87(+/- 0.5) degrees C. By combining the HeI photoelectron (PE) spectrum with the improved density function theory (DFT) calculations, we have concluded that the (CH3)3CO* radical has C3V symmetry and X2E ground state. The study does not only provide the ionization energies of different ionic states of the (CH3)3CO* radical for the first time, but also the first example in which there have been similar vibrational structures in different ionic states caused by removal of the electron on an orbital. It is also pointed out that (CH3)3CONO is a good source for obtaining the (CH3)3CO* radical beam, and that NO is a stable regent for the active radical. The results will promote the studies in electron spin resonance (ESR) research on the mechanisms of both the initiation of the formation of a new radical and the radical-chain polymerization in which the (CH3)3CO* radical participates.

Journal Article↗
PubMed2001

mu-Opioid receptor-mediated ERK activation involves calmodulin-dependent epidermal growth factor receptor transactivation.

Phosphorylation of the MAPK isoform ERK by G protein-coupled receptors involves multiple signaling pathways. One of these pathways entails growth factor receptor transactivation followed by ERK activation. This study demonstrates that a similar signaling pathway is used by the mu-opioid receptor (MOR) expressed in HEK293 cells and involves calmodulin (CaM). Stimulation of MOR resulted in both epidermal growth factor receptor (EGFR) and ERK phosphorylation. Data obtained with inhibitors of EGFR Tyr kinase and membrane metalloproteases support an intermediate role of EGFR activation, involving release of endogenous membrane-bound epidermal growth factor. Previous studies had demonstrated a role for CaM in opioid signaling based on direct CaM binding to MOR. To test whether CaM contributes to EGFR transactivation and ERK phosphorylation by MOR, we compared wild-type MOR with mutant K273A MOR, which binds CaM poorly, but couples normally to G proteins. Stimulation of K273A MOR with [D-Ala(2),MePhe(4),Gly-ol(5)]enkephalin (10-100 nm) resulted in significantly reduced ERK phosphorylation. Furthermore, wild-type MOR stimulated EGFR Tyr phosphorylation 3-fold more than K273A MOR, indicating that direct CaM-MOR interaction plays a key role in the transactivation process. Inhibitors of CaM and protein kinase C also attenuated [D-Ala(2),MePhe(4),Gly-ol(5)]enkephalin-induced EGFR transactivation in wild-type (but not mutant) MOR-expressing cells. This novel pathway of EGFR transactivation may be shared by other G protein-coupled receptors shown to interact with CaM.

Animals↗
PubMed2001

Single nucleotide polymorphisms in the human mu opioid receptor gene alter basal G protein coupling and calmodulin binding.

The mu opioid receptor (MOR) plays a central role in mediating acute and chronic effects of narcotic drugs. Three rare single nucleotide polymorphisms in the hMOR gene have been identified that cause amino acid substitutions in the third intracellular (i3) loop of MOR (R260H, R265H, and S268P). Genotyping 252 individuals of the Coriell collection identified one allele encoding the R265H-MOR variant and a new variant encoding D274N-MOR. Variants R260H-, R265H-, and S268P-MOR were constructed and transfected into HEK293 cells. Morphine stimulated G protein coupling of the three receptor variants to a maximal level approaching that of wild type MOR. In contrast, spontaneous, agonist-independent (basal) MOR signaling, proposed to play a role in opioid tolerance and dependence, was significantly reduced for R260H- and R265H-MOR. Moreover, domains within the i3 loop of MOR have been shown to interact with both G proteins and calmodulin (CaM). CaM binding was deficient for variants R265H- and S268P-MOR, suggesting that domains for G protein coupling and CaM binding overlap partially. Morphine pretreatment significantly enhanced basal G protein coupling of wild type MOR, which is thought to result from release of CaM. In contrast basal G protein coupling activity of the three variants was unaffected by morphine pretreatment consistent with diminished CaM regulation, low basal activity, or both. In conclusion, each of the three single nucleotide polymorphisms mapping to the i3 loop of MOR caused substantial changes in basal G protein coupling, CaM binding, or both. Carriers of the mutant alleles might display altered responses to narcotic analgesics.

Calmodulin↗
PubMed2001

Distribution and depression of the GABA(B) receptor in the spinal dorsal horn of adult rat.

gamma-Aminobutyric acid (GABA) is a principal inhibitory neurotransmitter in vertebrate nervous system. The metabotropic receptor for GABA, GABA(B) receptor, is characterized as a G protein-coupled receptor subtype. In the present study, GABA(B) receptor-like immunoreactivity (GABA(B)R-LI) in the rat spinal cord and dorsal root ganglion (DRG), as well as GABA(B) receptor-mediated depression in the spinal dorsal horn were examined by using immunohistochemistry and whole-cell voltage-clamp recording technique, respectively. Under light microscope, GABA(B)R-LI was densely found in laminae I and II of the dorsal horn. DRG cells of various diameters also showed GABA(B)R-LI. Electron microscopy further revealed that GABA(B)R-LI was also localized in terminals of myelinated, unmyelinated fibers as well as the somatodendritic sites of dorsal horn neurons. Bath application of a GABA(B) receptor agonist, baclofen (10 microM, 30 s), induced a slow outward (inhibitory) current in dorsal horn neurons. This slow current was depressed when the postsynaptic G protein-coupled receptor was inhibited, indicating the postsynaptic action of baclofen. Under the condition of postsynaptic GABA(B) receptor being inhibited, baclofen (10 microM, 60 s) depressed large (Abeta) and fine (C, Adelta) afferent fiber-evoked monosynaptic excitatory postsynaptic currents, indicating presynaptic inhibition of GABA(B) receptor on elicited neurotransmitter release. Taken together, the results suggest that baclofen-sensitive GABA(B) receptor is expressed pre- and postsynaptically on primary afferent fibers and neurons in the spinal dorsal horn; activation of GABA(B) receptor in the dorsal horn postsynaptically hyperpolarizes dorsal horn neurons and presynaptically inhibits primary afferents.

Age Factors↗
PubMed2001

Oscillatory electroosmosis-enhanced intra/inter-particle liquid transport and its primary applications in the preparative electrochromatography of proteins.

The concept of generating an oscillatory electroosmotic flux inside the porous particle to enhance the intra-particle mass transport was presented and a new kind of electrochromatography carried out in a five-compartment electrolyzer were developed. The adsorbent was packed in the central compartment, while the neighboring compartments were used as the elution compartments and the electrode compartments, respectively. Chromatographic separations of human serum albumin on Blue Sepharose Fast Flow, bovine serum albumin (BSA) on DEAE-Sepharose Fast Flow, and BSA on hydroxyapatite were carried out, respectively. The adsorption isotherms were shown to be independent of electric field, while the increase in the electric field strength resulted in a linear increase in the magnitude of electroosmotic flux and the improvement of the breakthrough behavior in all cases. The experiment results have demonstrated the effectiveness of the oscillatory electroosmosis in enhancing intra- and inter-particle mass transport and its high potential to large-scale chromatography.

Adsorption↗
PubMed2001

Activation of cardiac gene expression by myocardin, a transcriptional cofactor for serum response factor.

Serum response factor (SRF) regulates transcription of numerous muscle and growth factor-inducible genes. Because SRF is not muscle specific, it has been postulated to activate muscle genes by recruiting myogenic accessory factors. Using a bioinformatics-based screen for unknown cardiac-specific genes, we identified a novel and highly potent transcription factor, named myocardin, that is expressed in cardiac and smooth muscle cells. Myocardin belongs to the SAP domain family of nuclear proteins and activates cardiac muscle promoters by associating with SRF. Expression of a dominant negative mutant of myocardin in Xenopus embryos interferes with myocardial cell differentiation. Myocardin is the founding member of a class of muscle transcription factors and provides a mechanism whereby SRF can convey myogenic activity to cardiac muscle genes.

Amino Acid Sequence↗
PubMed2001

[Mutation analysis of tumor suppressor gene PTEN in bone and soft tissue tumors].

OBJECTIVE: To study tumor suppressor gene PTEN's role in the tumorigenesis of bone and soft tissue tumors. METHODS: Exon 4 approximately 9 of PTEN were amplified by polymerase chain reaction (PCR) for homozygous deletions in 110 tumor samples of patients with bone and soft tissue tumors, 3 tumor cell lines. PTEN mutations were detected by a combination of single strand conformation polymorphism (SSCP) analysis and DNA sequencing. Normal tissue beside tumors was used as control. RESULTS: There were always products of amplified exon 4 approximately 9 of PTEN, so no homozygous deletions existed. No homozygous deletions presented as variant band was not found by SSCP analysis. Two kinds of band type appeared in exon 8. A g/t polymorphism was found 32 bp from the splice donor site of intron 8 in six tumors. CONCLUSIONS: At the level of DNA, PTEN mutations do not play major roles in the tumorigenesis of bone and soft tissue tumors. The polymorphism of PTEN gene exists in Chinese people.

Asian People↗
PubMed2001

Alterations in the sensing and transport of phosphate and calcium by differentiating chondrocytes.

During endochondral bone formation and fracture healing, cells committed to chondrogenesis undergo a temporally restricted program of differentiation that is characterized by sequential changes in their phenotype and gene expression. This results in the manufacture, remodeling, and mineralization of a cartilage template on which bone is laid down. Articular chondrocytes undergo a similar but restricted differentiation program that does not proceed to mineralization, except in pathologic conditions such as osteoarthritis. The pathogenesis of disorders of cartilage development and metabolism, including osteochondrodysplasia, fracture non-union, and osteoarthritis remain poorly defined. We used the CFK2 model to examine the potential roles of phosphate and calcium ions in the regulatory pathways that mediate chondrogenesis and cartilage maturation. Differentiation was monitored over a 4-week period using a combination of morphological, biochemical, and molecular markers that have been characterized in vivo and in vitro. CFK2 cells expressed the type III sodium-dependent phosphate transporters Glvr-1 and Ram-1, as well as a calcium-sensing mechanism. Regulated expression and activity of Glvr-1 by extracellular phosphate and parathyroid hormone-related protein was restricted to an early stage of CFK2 differentiation, as evidenced by expression of type II collagen, proteoglycan, and Ihh. On the other hand, regulated expression and activity of a calcium-sensing receptor by extracellular calcium was most evident after 2 weeks of differentiation, concomitant with an increase in type X collagen expression, alkaline phosphatase activity and parathyroid hormone/parathyroid hormone-related protein receptor expression. On the basis of these temporally restricted changes in the sensing and transport of phosphate and calcium, we predict that extracellular phosphate plays a role in the commitment of chondrogenic cells to differentiation, whereas extracellular calcium plays a role at a later stage in their differentiation program.

Alkaline Phosphatase↗