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Biomedical subjects

Yan Sun

Publications and source records attributed to Yan Sun.

At least 181 records · Page 10Linked to original sources
PubMed2002

Gbeta gamma -independent constitutive association of Galpha s with SHP-1 and angiotensin II receptor AT2 is essential in AT2-mediated ITIM-independent activation of SHP-1.

Conventional mode of activation of SH2 domain-containing phosphatase 1 (SHP-1) by a single transmembrane (TM) inhibitory receptor such as killer cell inhibitory receptor, Fcgamma receptor type IIb1, and paired Ig-like receptors of inhibitory types requires tyrosine phosphorylation of immunoreceptor tyrosine-based inhibitory (ITIM) motifs in the cytoplasmic domains of the inhibitory receptors. Contrary to this paradigm, AT(2), a G protein-coupled 7TM receptor that does not undergo tyrosine phosphorylation in response to angiotensin II (Ang II) stimulation, also activates SHP-1. Here we show that SHP-1 constitutively and physically associates with AT(2) receptor in transfected COS-7 cells. On stimulation by Ang II, SHP-1 becomes activated and dissociated from AT(2) receptor, independent of pertussis toxin. Cotransfection of transducin G(betagamma) inhibits SHP-1/AT(2) association and the SHP-1 activation, whereas cotransfection of C-terminal of beta-adrenergic receptor kinase, which abrogates G(betagamma) signaling, facilitates SHP-1 activation. Surprisingly, SHP-1/AT(2) association and the SHP-1 activation requires the presence of G(alphas) as shown by differential coimmunoprecipitation, dominant negative G(alphas), constitutively active G(alphas), and G(alpha) peptides. A mutant AT(2) receptor D141A-R142L that is inactive in G(alpha) protein activation constitutively associates with SHP-1 and activates it. Together, these results indicate that G(alphas) alone, rather than exclusively in the form of G(alphabetagamma) heterotrimer may facilitate signal transduction for G protein-coupled receptors, suggesting a novel mechanism distinct from the classic paradigm of heterotrimeric G proteins. The AT(2)-mediated ITIM-independent activation of SHP-1 that is distinct from the conventional mode of activation, may represent a general paradigm for activation of SHP-1/2-class tyrosine phosphatases by G protein-coupled receptors.

Amino Acid Sequence↗
PubMed2002

Study on protein adsorption kinetics to a dye-ligand adsorbent by the pore diffusion model.

Adsorption kinetics of bovine serum albumin (BSA) and bovine hemoglobin (bHb) to Cibacron Blue 3GA (CB) modified Sepharose CL-6B has been studied. The effects of liquid-phase ionic strength and CB coupling density on the uptake rates of these two proteins in Tris-HCl buffer (pH 7.5) were evaluated by effective pore diffusivity derived from a pore diffusion model. The results showed that despite their similar molecular masses and sizes, the effects of aqueous-phase ionic strength and CB density on the effective pore diffusivities of BSA and bHb were distinctly different. The effective pore diffusivity of BSA to CB-Sepharose increased significantly with decreasing CB density and increasing liquid-phase ionic strength. This was considered due to the decrease in electrostatic repulsion between the BSA and CB molecules of like charge. That is, the increase in ionic strength and the decrease in CB coupling density reduced the electrostatic hindrance effect on BSA diffusion to CB-Sepharose, facilitating the hindered pore diffusion. In contrast, because of the higher isoelectric point of bHb (7.0) compared to BSA (4.7), bHb suffered little electrostatic hindrance effect during its diffusion to CB-Sepharose. Therefore, the effective pore diffusivity of bHb was unchanged with the change in liquid-phase ionic strength and CB coupling density.

Adsorption↗
PubMed2002

Analysis of diffusion models for protein adsorption to porous anion-exchange adsorbent.

The ion-exchange adsorption kinetics of bovine serum albumin (BSA) and gamma-globulin to an anion exchanger, DEAE Spherodex M, has been studied by batch adsorption experiments. Various diffusion models, that is, pore diffusion, surface diffusion, homogeneous diffusion and parallel diffusion models, are analyzed for their suitabilities to depict the adsorption kinetics. Protein diffusivities are estimated by matching the models with the experimental data. The dependence of the diffusivities on initial protein concentration is observed and discussed. The adsorption isotherm of BSA is nearly rectangular, so there is little surface diffusion. As a result, the surface and homogeneous diffusion models do not fit to the kinetic data of BSA adsorption. The adsorption isotherm of gamma-globulin is less favorable, and the surface diffusion contributes greatly to the mass transport. Consequently, both the surface and homogeneous diffusion models fit to the kinetic data of gamma-globulin well. The adsorption kinetics of BSA and gamma-globulin can be very well fitted by parallel diffusion model, because the model reflects correctly the intraparticle mass transfer mechanism. In addition, for both the favorably bound proteins, the pore diffusion model fits the adsorption kinetics reasonably well. The results here indicate that the pore diffusion model can be used as a good approximate to depict protein adsorption kinetics for protein adsorption systems from rectangular to linear isotherms.

Adsorption↗
PubMed2002

Steric mass-action model for dye-ligand affinity adsorption of protein.

The steric mass-action (SMA) model has been reported in the literature for ion-exchange and metal-affinity interaction adsorption equilibrium of proteins. In this work, an SMA model was developed for protein adsorption equilibrium to dye-ligand affinity adsorbent, Cibacron Blue-modified Sepharose CL-6B (CB-Sepharose). Static adsorption experiments with bovine serum albumin as a model protein were carried out to determine the model parameters, that is, equilibrium constant (K), characteristic number of binding sites (n), and steric factor (sigma). It was found that the linear model parameters, K and n decreased with the increase of ionic strength, while the nonlinear parameter, sigma, increased with ionic strength and the dye-ligand concentration. Thus, expressions correlating these parameters with the dye-ligand concentration and/or ionic strength were derived. With these correlations, the SMA model gave promising results in predicting protein adsorption isotherms. Thus, it is considered that the model would be useful in the theoretical analysis of dye-ligand affinity chromatography.

Adsorption↗
PubMed2002

Rac function and regulation during Drosophila development.

Rac GTPases regulate the actin cytoskeleton to control changes in cell shape. To date, the analysis of Rac function during development has relied heavily on the use of dominant mutant isoforms. Here, we use loss-of-function mutations to show that the three Drosophila Rac genes, Rac1, Rac2 and Mtl, have overlapping functions in the control of epithelial morphogenesis, myoblast fusion, and axon growth and guidance. They are not required for the establishment of planar cell polarity, as had been suggested on the basis of studies using dominant mutant isoforms. The guanine nucleotide exchange factor, Trio, is essential for Rac function in axon growth and guidance, but not for epithelial morphogenesis or myoblast fusion. Different Rac activators thus act in different developmental processes. The specific cellular response to Rac activation may be determined more by the upstream activator than the specific Rac protein involved.

Animals↗
PubMed2002

Rac GTPases control axon growth, guidance and branching.

Growth, guidance and branching of axons are all essential processes for the precise wiring of the nervous system. Rho family GTPases transduce extracellular signals to regulate the actin cytoskeleton. In particular, Rac has been implicated in axon growth and guidance. Here we analyse the loss-of-function phenotypes of three Rac GTPases in Drosophila mushroom body neurons. We show that progressive loss of combined Rac1, Rac2 and Mtl activity leads first to defects in axon branching, then guidance, and finally growth. Expression of a Rac1 effector domain mutant that does not bind Pak rescues growth, partially rescues guidance, but does not rescue branching defects of Rac mutant neurons. Mosaic analysis reveals both cell autonomous and non-autonomous functions for Rac GTPases, the latter manifesting itself as a strong community effect in axon guidance and branching. These results demonstrate the central role of Rac GTPases in multiple aspects of axon development in vivo, and suggest that axon growth, guidance and branching could be controlled by differential activation of Rac signalling pathways.

Amino Acid Sequence↗
PubMed2002

Fabrication and characterization of a rigid magnetic matrix for protein adsorption.

This article describes the fabrication and characterization of a novel magnetic poly(glycidyl methacrylate-triallyl isocyanurate-divinylbenzene) matrix containing magnetite colloids. The results showed that the matrix was superparamagnetic and could be separated magnetically from a suspension in a few seconds. Protein adsorption properties of diethylamine-derivatized matrix were characterized with bovine serum albumin (BSA) as a model protein. The static capacity determined by batch adsorption was 79 mg/ml wet matrix. Kinetic study gave an effective diffusivity of BSA of 5.0 x 10(-13) m2/s in the matrix at an initial BSA concentration in the liquid phase of 1.0 mg/ml. Stability of the matrix was confirmed by recycling of the matrix in protein adsorptions.

Adsorption↗
PubMed2002

Nd-Fe-B alloy-densified agarose gel for expanded bed adsorption of proteins.

Novel dense composite adsorbents for expanded bed adsorption of protein have been fabricated by coating 4% agarose gel onto Nd-Fe-B alloy powder by a water-in-oil emulsification method. Two composite matrices, namely Nd-Fe-B alloy-densified agarose (NFBA) gels with different size distributions and densities, NFBA-S (50-165 microm, 1.88 g/ml) and NFBA-L (140-300 microm, 2.04 g/ml), were produced. Lysozyme was used as a model protein to test the adsorption capacity and kinetics for the NFBA gels modified by Cibacron blue 3GA (CB-NFBA gels). Liquid-phase dispersion behavior in the expanded beds was examined by measurements of residence time distributions, and compared with that of Streamline SP (Amersham-Pharmacia Biotech, Sweden). The dependence of axial mixing in the expanded beds on flow velocity, bed expansion degree. settled bed height, and viscosity of liquid phase was investigated. Breakthrough curves of lysozyme in the expanded beds of the CB-NFBA gels were also examined. The dynamic binding capacity at 5% breakthrough was 23.3 mg/ml matrix for the CB-NFBA-S gels, and 16.7 mg/ml matrix for the CB-NFBA-L, at a flow velocity of 220 cm/h. The results indicate that the NFBA gels are promising for expanded bed adsorption of proteins.

Adsorption↗
PubMed2002

ECG signal conditioning by morphological filtering.

Clinically obtained electrocardiographic (ECG) signals are often contaminated with different types of noise and baseline drifting commonly occurs. In order to facilitate automated ECG analysis, signal conditioning is undoubtedly a necessity. In this paper, a modified morphological filtering (MMF) technique is used for signal conditioning in order to accomplish baseline correction and noise suppression with minimum signal distortion. Compared with existing methods for ECG signal conditioning, MMF performs well in terms of the filtering characteristics, low signal distortion ratio, low computational burden as well as good noise suppression ratio and baseline correction ratio.

Algorithms↗
PubMed2002

Effect of doxazosin on stretch-activated adenosine triphosphate release in bladder urothelial cells from patients with benign prostatic hyperplasia.

OBJECTIVES: Recent data suggest that the bladder urothelium may have a sensory function by way of release of adenosine triphosphate (ATP) during stretch, which then acts as a sensory neurotransmitter. Because benign prostatic hyperplasia (BPH) can give rise to irritative (hypersensory) voiding patterns, we questioned whether the bladder urothelium from patients with BPH released more ATP during in vitro stretch and whether doxazosin, an alpha(1)-adrenoceptor blocker, affects this purinergic mechanism. METHODS: Bladder urothelial biopsies from patients with BPH (n = 4) and controls (n = 4) were cultured using established techniques. In vitro stretch was performed with a Flexcell 2000 device that uses vacuum to deform the cell growth surface to impart a stretch force. Doxazosin (5 microM and 20 microM) was added to cells, and supernatants were collected at various points for ATP assay. ATP was assayed using the luciferin-luciferase reaction. ATP data were normalized to the time 0 value and expressed as a percentage of the baseline value. RESULTS: After 96 hours of stretch, the BPH urothelial cells released significantly more ATP than did the control urothelial cells (62.6% +/- 11.2% versus 24.2% +/- 5.4%, P = 0.005) and nonstretched BPH urothelial cells (62.6% +/- 11.2% versus 15.1% +/- 5.1%, P = 0.004). The augmented release of ATP by stretched BPH bladder urothelial cells was completely blocked by treatment with 20 microM doxazosin. CONCLUSIONS: Irritative voiding secondary to BPH may arise from increased ATP release by bladder urothelium during stretch. Doxazosin inhibits ATP release by way of an unknown mechanism that may or may not involve the alpha1-adrenoreceptor. Treatment for hypersensory voiding symptoms secondary to BPH might also target the urothelial purinergic pathway.

Adenosine Triphosphate↗
PubMed2002

Both metal binding sites in the homodimer are required for metalloregulation by the CadC repressor.

The cadCA operon of plasmid pI258, which confers resistance to the soft metals Cd(II), Pb(II) and Zn(II), is regulated by CadC, a metal-responsive transcriptional repressor. CadC is a 27.6 kDa homodimer composed of two 122-residue monomers. Three cysteine residues, Cys-7, Cys-58 and Cys-60, have been shown to be required for sensing soft metals. Thus, the repressor has two potential inducer binding sites, one on each monomer. However, it is not known whether both binding sites are required for derepression or whether binding of metal to a single site would result in transcript. In this study, heterodimers were purified in which one binding site was wild type and the other had substitutions of the cysteine residues. The wild type-mutant heterodimers retained the ability to bind to cad operator/promoter DNA but did not dissociate from the DNA upon addition of soft metal ions. The results indicate that both subunits in the dimer must have functional metal binding sites for metal sensing to lead to derepression

Bacterial Proteins↗
PubMed2002

A retrospective look at the cationic peanut peroxidase structure.

The cationic peanut peroxidase has been studied in detail, not only with regard to its peptide structure, but also to the sites and role of the three moieties linked to it. Peanut peroxidase lends itself well to a close examination as a potential example for other plant peroxidase studies. It was the first plant peroxidase for which a 3-D structure was derived from crystals, with the glycans intact. Subsequent analysis of peroxidases structures from other plants have not shown great differences to that of the peanut peroxidase. As the period of proteomics follows on the era of genomics, the study of glycans has been brought back into focus. With the potential use of peroxidase as a polymerization agent for industry, there are some aspects of the overall structure that should be kept in mind for successful use of this enzyme. A variety of techniques are now available to assay for these structures/moieties and their roles. Peanut peroxidase data are reviewed in that light, as well as defining some true terms for isozymes. Because a high return of the enzyme in a pure form has been obtained from cultured cells in suspension culture, a brief review of this is also offered.

Arachis↗
PubMed2002

Molecular characterization of the TCP11 gene which is the human homologue of the mouse gene encoding the receptor of fertilization promoting peptide.

A human testis-specific gene was isolated by subtractive hybridization between the cDNA pools of adult and fetal testes, followed by rapid amplification of cDNA ends (RACE). This gene sequence is highly homologous to a large portion of the mouse Tcp11 gene which is important in sperm function because it encodes the receptor for fertilization-promoting peptide (FPP). The gene was mapped to human chromosome band 6p21 by fluorescence in-situ hybridization. The 9 exon gene spans a 22.8 kp genomic DNA sequence. The mature processed message encodes a 441 amino acid protein that is highly homologous to the mouse 566 amino acid protein after the first 142 amino acids. Results of Northern blot and RT-PCR analyses of RNA extracted from human tissues revealed that the gene is only expressed in fertile adult testes, but not in azoospermic testes, fetal testes nor in other human tissues. Taken together, our results along with the mouse Tcp11 function suggest that TCP11 gene is important in sperm function and fertility.

Amino Acid Sequence↗
PubMed2002

Cryopreserved cell monolayers for rapid detection of herpes simplex virus and influenza virus.

Cryopreserved cell monolayers are a new cell culture technology intended to ensure the availability of cells in the laboratory for virus detection. Two cryopreserved cell monolayers, ELVIS for the detection of herpes simplex virus (HSV) and R-Mix for the detection of influenza virus, were evaluated. The results indicated that fresh and cryopreserved cell monolayers are comparable in sensitivity for the detection of HSV and influenza virus. The cells retain the same level of sensitivity for up to 4 months at -80 degrees C.

Animals↗
PubMed2002

Engineered BGMK cells for sensitive and rapid detection of enteroviruses.

Decay-accelerating factor (DAF) has been reported to be a cellular receptor for several enteroviruses. Buffalo green monkey kidney (BGMK) cells expressing human DAF (BGMK-hDAF cells) showed increased susceptibility and sensitivity to several types of enteroviruses compared to wild-type BGMK cells. When 17 frozen positive clinical samples were tested, BGMK cells detected 8 and BGMK-hDAF cells detected 16. Since the CaCo-2 cell line has been documented to support the replication of most enteroviruses, CaCo-2 cells were mixed with BGMK-hDAF cells in order to increase the number of viruses detected. Thirty-four frozen clinical samples that previously had tested positive for enteroviruses were tested, and the following numbers were detected: 33 of 34 by CaCo-2/BGMK-hDAF cells, 29 of 34 by CaCo-2/BGMK cells, 28 of 34 by H292/RD (E-mix A) and A-549/BGMK (E-mix B) cells, and 26 of 34 by MRC-5 and pRhMK cells.

Animals↗
PubMed2002

[Polymorphisms of four microsatellite markers tightly linked with polycystic kidney disease 2 gene in Chinese].

OBJECTIVE: To investigate the polymorphisms of four microsatellites, D4S1534, D4S1563, D4S423 and D4S414, which are tightly linked to polycystic kidney disease 2 (PKD2) gene, and hence to provide a basis for studying the heterogeneity of adult polycystic kidney disease (APKD). METHODS: An analysis on the DNA of some unrelated Chinese people was performed using polymerase chain reaction (PCR), polyacrylamide gel electrophoresis (PAGE) and silver staining. RESULTS: In Chinese Hans, there are 11 alleles of D4S1534, and their sizes are 142-162 bp 14 alleles of D4S1563, 205-235 bp 17 alleles of D4S423, 103-135 bp; and 15 alleles of D4S414, 236-264 bp. The polymorphism information contents of the four microsatellites are 0.872, 0.844, 0.921 and 0.871 respectively. CONCLUSION: In the Chinese Han people studied above, the four microsatellite markers that have many alleles are highly polymorphic genetic markers and may serve as the data of population genetics, suggesting that all four microsatellites could be used in studies on heterogeneity of APKD, linkage analysis of APKD and forensic personal identification.

Alleles↗