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D Tang

Publications and source records attributed to D Tang.

At least 73 records · Page 4Linked to original sources
PubMed1999

Towards rice genome scanning by map-based AFLP fingerprinting.

Map-based DNA fingerprinting with AFLP markers provides a fast method for scanning the rice genome. Three hundred AFLP markers identified with ten primer combinations were mapped in two rice populations. The genetic maps were aligned and almost full coverage of the rice genome was obtained. The transferability of AFLP markers between indica x japonica and indica x indica crosses was tested. The chromosomes were divided into DNA Fingerprint Linkage Blocks (DFLBs) defined by specific AFLP markers. Using these blocks, the degree of similarity or divergence within specific chromosome regions was calculated for nine varieties. Applications of map-based fingerprinting for biodiversity studies and maker-assisted selection are discussed.

Chromosome Mapping↗
PubMed1999

Membrane lipid alpha-crystallin interaction and membrane Ca2+ -ATPase activities.

PURPOSE: To determine the effect of alpha-crystallin binding on lens membrane lipid characteristics and the stability of Ca2+ -ATPase activity when challenged with H2O2 or elevated temperatures. METHODS: Alpha-Crystallin binding to muscle sarcoplasmic reticulum membranes was quantified using a centrifugation protocol. Alpha-Crystallin binding to lens epithelial lipids was measured by a fluorescence energy transfer technique. Lipid phase transition temperature and lipid order was measured using fluorescence spectroscopy. Ca2+ -ATPase activity was measured using classical biochemical assays. RESULTS: The main phase transition temperatures of multilamellar vesicles composed of sphingomyelin or lipids extracted from bovine lens were 40 degrees C and 20 degrees C, respectively. In the presence of saturating amounts of alpha-crystallin, the phase transition temperature and lipid order of both sphingomyelin and lens lipid membranes remained almost the same as that without alpha-crystallin. The interaction of alpha-crystallin and lipid is likely to be restricted to the membrane surface. The binding of alpha-crystallin did not influence the oxidative or thermal inactivation of the Ca2+ -ATPase pump. CONCLUSION: Alpha-Crystallin-lens membrane binding does not protect the Ca2+ -ATPase pump from thermal derangement or oxidation by H2O2.

Animals↗
PubMed1999

Malaria prophylaxis using azithromycin: a double-blind, placebo-controlled trial in Irian Jaya, Indonesia.

New drugs are needed for preventing drug-resistant Plasmodium falciparum malaria. The prophylactic efficacy of azithromycin against P. falciparum in malaria-immune Kenyans was 83%. We conducted a double-blind, placebo-controlled trial to determine the prophylactic efficacy of azithromycin against multidrug-resistant P. falciparum malaria and chloroquine-resistant Plasmodium vivax malaria in Indonesian adults with limited immunity. After radical cure therapy, 300 randomized subjects received azithromycin (148 subjects, 750-mg loading dose followed by 250 mg/d), placebo (77), or doxycycline (75, 100 mg/d). The end point was slide-proven parasitemia. There were 58 P. falciparum and 29 P. vivax prophylaxis failures over 20 weeks. Using incidence rates, the protective efficacy of azithromycin relative to placebo was 71.6% (95% confidence interval [CI], 50.3-83.8) against P. falciparum malaria and 98.9% (95% CI, 93.1-99.9) against P. vivax malaria. Corresponding figures for doxycycline were 96.3% (95% CI, 85.4-99.6) and 98% (95% CI, 88.0-99.9), respectively. Daily azithromycin offered excellent protection against P. vivax malaria but modest protection against P. falciparum malaria.

Adolescent↗
PubMed1999

A nonlinear axisymmetric model with fluid-wall interactions for steady viscous flow in stenotic elastic tubes.

Arteries with high-grade stenoses may compress under physiologic conditions due to negative transmural pressure caused by high-velocity flow passing through the stenoses. To quantify the compressive conditions near the stenosis, a nonlinear axisymmetric model with fluid-wall interactions is introduced to simulate the viscous flow in a compliant stenotic tube. The nonlinear elastic properties of the tube (tube law) are measured experimentally and used in the model. The model is solved using ADINA (Automatic Dynamic Incremental Nonlinear Analysis), which is a finite element package capable of solving problems with fluid-structure interactions. Our results indicate that severe stenoses cause critical flow conditions such as negative pressure and high and low shear stresses, which may be related to artery compression, plaque cap rupture, platelet activation, and thrombus formation. The pressure filed near a stenosis has a complex pattern not seen in one-dimensional models. Negative transmural pressure as low as -24 mmHg for a 78 percent stenosis by diameter is observed at the throat of the stenosis for a downstream pressure of 30 mmHg. Maximum shear stress as a high as 1860 dyn/cm2 occurs at the throat of the stenoses, while low shear stress with reversed direction is observed right distal to the stenosis. Compressive stresses are observed inside the tube wall. The maximal principal stress and hoop stress in the 78 percent stenosis are 80 percent higher than that from the 50 percent stenosis used in our simulation. Flow rates under different pressure drop conditions are calculated and compared with experimental measurements and reasonable agreement is found for the prebuckling stage.

Animals↗
PubMed1999

Mechanosensitive tyrosine phosphorylation of paxillin and focal adhesion kinase in tracheal smooth muscle.

We investigated the role of the integrin-associated proteins focal adhesion kinase (FAK) and paxillin as mediators of mechanosensitive signal transduction in tracheal smooth muscle. In muscle strips contracted isometrically with ACh, we observed higher levels of tyrosine phosphorylation of FAK and paxillin at the optimal muscle length (Lo) than at shorter muscle lengths of 0.5 or 0.75 Lo. Paxillin phosphorylation was also length sensitive in muscles activated by K+ depolarization and adjusted rapidly to changes in muscle length imposed after contractile activation by either ACh or K+ depolarization. Ca2+ depletion did not affect the length sensitivity of paxillin and FAK phosphorylation in muscles activated with ACh, indicating that the mechanotransduction process can be mediated by a Ca2+-independent pathway. Since Ca2+-depleted muscles do not generate significant active tension, this suggests that the mechanotransduction mechanism is sensitive to muscle length rather than tension. We conclude that FAK and paxillin participate in an integrin-mediated mechanotransduction process in tracheal smooth muscle. We propose that this pathway may initiate alterations in smooth muscle cell structure and contractility via the remodeling of actin filaments and/or via the mechanosensitive regulation of signaling molecules involved in contractile protein activation.

Acetylcholine↗
PubMed1999

Alpha-crystallin/lens lipid interactions using resonance energy transfer.

Resonance energy transfer was used to study the interaction of alpha-crystallin with lens cortex lipid vesicles. The binding of alpha-crystallin to cortex lipid vesicles and the preincubation temperature dependence of the binding were confirmed. In this study, the tryptophan of alpha-crystallin was used as the energy donor, and the fluorescence probe N-(5-dimethylaminonaphthalene-1-sulfonyl)-1, 2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (dansyl DHPE) was chosen as the energy acceptor. Lens cortex lipid vesicles were preincorporated with dansyl DHPE. Energy transfer from the tryptophan of alpha-crystallin to dansyl DHPE was found and the energy transfer efficiency was calculated. There was a higher energy transfer efficiency between alpha-crystallin and dansyl DHPE when alpha-crystallin was preincubated at 65 degrees C compared to 22 degrees C. Data confirmed the binding of alpha-crystallin to lens cortex lipid and showed that alpha-crystallin bound more closely to the surface of cortex vesicles when it was preincubated at a higher temperature. This is probably due to the exposure of hydrophobic surfaces when alpha-crystallin is preincubated at a higher temperature.

Absorption↗
PubMed1999

Molecular and genetic damage from environmental tobacco smoke in young children.

To assess the risks of early life exposure to environmental tobacco smoke (ETS), we tested whether four biomarkers in peripheral blood were associated with home ETS exposure in Hispanic and African-American children. The biomarkers included cotinine (a metabolite of nicotine) and three indicators of molecular and genetic damage from mutagens/carcinogens, protein adducts formed by the carcinogens 4-aminobiphenyl (4-ABP) and polycyclic aromatic hydrocarbons (PAHs), and sister chromatid exchanges (SCEs). We also explored possible ethnic differences in biomarkers. The study cohort comprised 109 Hispanic and African-American preschool children (1-6 years of age). Plasma cotinine was analyzed by gas chromatography, 4-ABP-hemoglobin adducts by gas chromatography-mass spectroscopy, PAH-albumin adducts by ELISA, and SCEs by cytogenetic techniques. Data on the amount of smoking by mothers (average 10.5 cigarettes per day) and other household members and regular visitors (average 6.5 cigarettes per day) were obtained by interview-administered questionnaires. Cotinine, 4-ABP-hemoglobin adducts, and PAH-albumin were significantly higher (P < 0.05) in the ETS-exposed children compared with the unexposed. SCEs were marginally higher (P = 0.076). African-American children had higher levels of cotinine (P = 0.059) and PAH-albumin (P = 0.02) than Hispanic children, after controlling for exposure to ETS. These results indicate molecular and genetic damage in minority children with

Aminobiphenyl Compounds↗
PubMed1999

[Evaluation of biliary CEA in the diagnosis of colorectal cancer with liver metastases].

OBJECTIVE: To evaluate the clinical significance of biliary CEA for diagnosing colorectal cancer with liver metastases. METHODS: Three groups of patients were examined: 30 patients with benign disease in group A, 30 patients with colorectal cancer in group B, and 15 colorectal cancer patients with liver metastasis in group C. Fasting venous blood was drawn, and in the meatime, bile was collected by duodenal tube drainage before operation. CEA was detected with immunosorbent assay kit supplied by Boehringer. RESULTS: The serum level of CEA in group A, B, and C was (3.2 +/- 0.5) ng/ml, (11.7 +/- 2.4) ng/ml, and (14.8 +/- 3.5) ng/ml, respectively. The biliary CEA level was (23.7 +/- 8.6) ng/ml, (129.4 +/- 36) ng/ml, and (862 +/- 227) ng/ml, respectively. The difference in biliary CEA between group B and group C was statistically significant (P < 0.01). CONCLUSION: CEA level in the bile collected by duodenal drainage helps confirm liver metastasis in patients with colorectal cancer.

Adult↗
PubMed1998

Cleavage of DFF-45/ICAD by multiple caspases is essential for its function during apoptosis.

Apoptosis involves the proteolysis of specific cellular proteins by a group of cysteine proteases known as caspases. Many of these cellular targets are either functionally inactivated (e.g. poly(ADP-ribose) polymerase) or activated (e.g. other caspases, gelsolin) by such processing, thereby facilitating the cell death process. Caspase 3 is involved in the processing of many of these proteins. Recently, however, it was reported that caspase 3 is dispensable for the cleavage of a large number of cellular caspase substrates during apoptosis. Among these substrates is DFF-45/ICAD, a subunit of the heterodimeric DNA fragmentation factor (DFF), otherwise known as caspase-activated DNase (CAD), that mediates genomic DNA degradation during apoptosis. Conversely, others have reported that caspase 3 is essential for the cleavage and activation of DFF-45/ICAD. To resolve this controversy we examined DFF-45/ICAD processing during apoptosis in MCF-7 breast carcinoma cells that lack functional caspase 3 and in MCF-7 cells expressing caspase 3. We found that DFF-45/ICAD is cleaved by two distinct caspases, one of which is caspase 3. Furthermore, cleavage of the carboxyl-terminal region of DFF-45/ICAD, which is necessary for activation of the enzyme, requires functional caspase 3. In the absence of caspase 3 cleavage of the amino-terminal region of DFF-45/ICAD by another caspase occurs, but the DFF-45 enzyme remains inactive.

Apoptosis↗
PubMed1998

A statistical paradigm for neural spike train decoding applied to position prediction from ensemble firing patterns of rat hippocampal place cells.

The problem of predicting the position of a freely foraging rat based on the ensemble firing patterns of place cells recorded from the CA1 region of its hippocampus is used to develop a two-stage statistical paradigm for neural spike train decoding. In the first, or encoding stage, place cell spiking activity is modeled as an inhomogeneous Poisson process whose instantaneous rate is a function of the animal's position in space and phase of its theta rhythm. The animal's path is modeled as a Gaussian random walk. In the second, or decoding stage, a Bayesian statistical paradigm is used to derive a nonlinear recursive causal filter algorithm for predicting the position of the animal from the place cell ensemble firing patterns. The algebra of the decoding algorithm defines an explicit map of the discrete spike trains into the position prediction. The confidence regions for the position predictions quantify spike train information in terms of the most probable locations of the animal given the ensemble firing pattern. Under our inhomogeneous Poisson model position was a three to five times stronger modulator of the place cell spiking activity than theta phase in an open circular environment. For animal 1 (2) the median decoding error based on 34 (33) place cells recorded during 10 min of foraging was 8.0 (7.7) cm. Our statistical paradigm provides a reliable approach for quantifying the spatial information in the ensemble place cell firing patterns and defines a generally applicable framework for studying information encoding in neural systems.

Action Potentials↗
PubMed1998

The granulin/epithelin precursor abrogates the requirement for the insulin-like growth factor 1 receptor for growth in vitro.

3T3 cells null for the type 1 insulin-like growth factor receptor are refractory to stimulation by a variety of purified growth factors that are known to be required for the stimulation of other 3T3 cells. However, these cells, known as R- cells, grow in serum-supplemented medium and also in media conditioned by certain cell lines. We report here the purification of a growth factor that stimulates DNA synthesis (and growth) of R- cells. The growth factor, purified to homogeneity by SDS-polyacrylamide gel electrophoresis, was identified as the granulin/epithelin precursor by an accurate determination of the masses of endoproteinase Lys-C peptides using matrix-assisted laser desorption ionization mass spectrometry, followed by a data base search. The granulin/epithelin precursor is a little known growth factor, secreted by a variety of epithelial and hemopoietic cells. It is at present the only purified growth factor that can stimulate the growth of mouse embryo fibroblasts null for the type 1 insulin-like growth factor receptor.

3T3 Cells↗
PubMed1998

Phosphorylation of PITSLRE p110 isoforms accompanies their processing by caspases during Fas-mediated cell death.

A number of cellular proteins have been identified as caspase targets during cell death, including the PITSLRE protein kinases. These targets generally fall into one of three possible categories: 1) other caspases, 2) proteins that are inactivated during apoptosis, and 3) proteins that are required for execution of the cell death program. However, not all proteins are cleaved by caspases during apoptosis. Why only specific proteins are destined to be processed by caspases during cell death is currently not clear. Here we show that multiple caspase-like activities are involved in the processing of the PITSLRE p110 isoforms during Fas-induced apoptosis in Jurkat T-cells. Three p110 caspase cleavage sites have been mapped to the amino-terminal domain of p110 and verified by site-directed mutagenesis. Curiously, the mutagenesis studies revealed that cleavage of two juxtaposed caspase sites is necessary for the complete processing of this protein during cell death in vivo. Finally, we demonstrate that the PITSLRE p110 protein is rapidly phosphorylated during Fas-induced apoptosis in Jurkat cells and that phosphorylation of an amino-terminal portion of the protein may enhance caspase cleavage in this region.

Apoptosis↗
PubMed1998

Lipid interactions with human antiphospholipid antibody, beta 2-glycoprotein 1, and normal human IgG using the fluorescent probes NBD-PE and DPH.

Recurrent venous thrombosis, arterial thrombosis and pregnancy losses are clinical manifestation associated with antiphospholipid antibody (aPL) that recognizes negatively charged phospholipid antigens. Enzyme-linked immunosorbent assays (ELISA) are generally used to determine the presence and specificity of aPL. In this paper, a fluorescence spectroscopy method has been applied, through monitoring the alteration of fluorescence intensity and anisotropy of a fluorophore that was incorporated in liposomes to explore the changes of molecular structure or configuration elicited by the binding aPL with phospholipid antigens. The bilayer surface was markedly ordered by aPL binding as indicated by the surface-sensitive probe NBD-PE. The binding of aPL on the bilayer surface is saturable. The saturation concentration of aPL is 40% (w/w, aPL/lipid) for cardiolipin membranes. The binding of aPL on cardiolipin took place in the absence of beta 2-GP1. The addition of beta 2-GP1 further increased the anisotropy and decreased the intensity of fluorescence. The binding of aPL is predominantly attributed to electrostatic interaction, but the configuration of the acyl chains of phospholipid also plays a role. It is found that the thermal history is important for aPL binding. The incubation at 37 degrees C is more favorable for aPL binding than ambient temperature. Normal human serine (IgG-NHS) did not elicit any distinct change of NBD-PE fluorescence, which indicates it does not interact with the lipid.

Animals↗
PubMed1998

The molecular basis of viral oncolysis: usurpation of the Ras signaling pathway by reovirus.

NIH-3T3 cells, which are resistant to reovirus infection, became susceptible when transformed with activated Sos or Ras. Restriction of reovirus proliferation in untransformed NIH-3T3 cells was not at the level of viral gene transcription, but rather at the level of viral protein synthesis. An analysis of cell lysates revealed that a 65 kDa protein was phosphorylated in untransformed NIH-3T3 cells, but only after infection with reovirus. This protein was not phosphorylated in infected or uninfected transformed cells. The 65 kDa protein was determined to be the double-stranded RNA-activated protein kinase (PKR), whose phosphorylation leads to translation inhibition. Inhibition of PKR phosphorylation by 2-aminopurine, or deletion of the Pkr gene, led to drastic enhancement of reovirus protein synthesis in untransformed cells. The emerging picture is one in which early viral transcripts trigger PKR phosphorylation in untransformed cells, which in turn leads to inhibition of translation of viral genes; this phosphorylation event is blocked by an element(s) in the Ras pathway in the transformed cells, allowing viral protein synthesis to ensue. The usurpation of the Ras signaling pathway therefore constitutes the basis of reovirus oncolysis.

3T3 Cells↗
PubMed1998

Association of neurofilament proteins with neuronal Cdk5 activator.

Cdk5 exists in brain extracts in multiple forms, one of which is a macromolecular protein complex comprising Cdk5, neuron-specific Cdk5 activator p35nck5a and other protein components (Lee, K.-Y., Rosales, J. L., Tang, D., and Wang, J.H. (1996) J. Biol. Chem. 271, 1538-1543). The yeast two-hybrid system was employed to identify p35nck5a-interacting proteins from a human brain cDNA library. One of the isolated clones encodes a fragment of glial fibrillary acidic protein, which is a glial-specific protein. Sequence alignment revealed significant homology between the p35nck5a-binding fragment of glial fibrillary acidic protein and corresponding regions in neurofilaments. The association between p35nck5a and neurofilament medium molecular weight subunit (NF-M) was confirmed by both the yeast two-hybrid assay and direct binding of the bacteria-expressed proteins. The p35nck5a binding site on NF-M was mapped to a carboxyl-terminal region of the rod domain, in close proximity to the putative Cdk5 phosphorylation sites in NF-M. A region immediately amino-terminal to the kinase-activating domain in p35nck5a is required for its binding with NF-M. In in vitro binding assays, NF-M binds both monomeric p35nck5a and the Cdk5/p35nck5a complex. The binding of NF-M has no effect on the kinase activity of Cdk5/p35nck5a.

Amino Acid Sequence↗
PubMed1998

Influence of cholesterol on the interaction of alpha-crystallin with phospholipids.

The influence of cholesterol on the binding of alpha-crystallin to pure phospholipid membranes was studied. The rationale of this investigation stems from two unique aspects of human lens cells: an unusually high level of cholesterol in the membranes and the specific binding of alpha-crystallin to membranes. In the absence of cholesterol, binding of alpha-crystallin liposomes composed of either sphingomyelin, disteroyl-phosphatidylcholine or egg-phosphatidylcholine caused a decrease in the fluorescence intensity and anisotropy of the fluorophore NBD-PE. Since this fluorescence probe resides in the polar headgroup region of the membrane, the observed changes indicated that the binding of alpha-crystallin affected the structure of these membrane regions. The ability of alpha-crystallin to modulate membrane structure suggests yet another potential role for this lens protein. Addition of cholesterol markedly decreased the binding of alpha-crystallin to liposomes composed of either sphingomyelin or disteroylphosphatidylcholine and antagonized the capacity of bound alpha-crystallin to decrease membrane surface order. This antagonism could be explained by the ability of cholesterol to directly decrease the anisotropy of the fluorophore in sphingomyelin membranes unexposed to alpha-crystallin. Thus, with cholesterol present, a further decrease in membrane order upon subsequent binding of alpha-crystallin was less likely. The results obtained with the sphingomyelin liposomes are considered most meaningful, since sphingomyelins are the principal phospholipids in the human lens nuclear membrane and cholesterol preferentially interacts with sphingomyelin. We conclude that cholesterol in lipid membranes can antagonize the binding of alpha-crystallin and thus interfere with the capacity of bound alpha-crystallin to alter membrane order. We suggest that such actions of cholesterol might serve to preserve lens membrane structure in the physiological state where the concentration of soluble alpha-crystallin is great.

Animals↗
PubMed1998

Temperature induced structural changes of beta-crystallin and sphingomyelin binding.

The study of the binding of alpha-crystallin to membranes is potentially important for understanding the function of alpha-crystallin in the ocular lens and the formation of cataracts. Using fluorescence probes, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3 -phosphoethanolamine, triethylammonium salt (NBD-PE) and (1,1'-bi(4-anilino)naphthalene-5,5'-disulfonic acid, dipotassium salt (bis-ANS), the temperature dependence of the binding of alpha-crystallin to sphingomyelin liposomes, and the structural changes of alpha-crystallin and sphingomyelin induced by temperature were studied. The influence of the binding of alpha-crystallin on the mobility of the head group region of liposomes of sphingomyelin was dependent on the thermal history of alpha-crystallin. Binding of alpha-crystallin to sphingomyelin caused a decrease in the anisotropy of the fluorophore NBD-PE at or below 37 degrees C. However, when alpha-crystallin or the mixture of alpha-crystallin/sphingomyelin were preincubated near the secondary structure phase transition temperature of 60 degrees C, an increase of the anisotropy of NBD-PE (decrease of lipid head group mobility) was observed when measured at 22 degrees C or 37 degrees C. An inflection near 47 degrees C in the curve of fluorescence anisotropy of bis-ANS pre-incorporated into the alpha-crystallin corresponded to a 3 degrees or 4 degrees structural change of alpha-crystallin. alpha-Crystallin either increases or decreases the flexibility of the head group of sphingomyelin liposomes depending on its structure.

Animals↗