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Biomedical subjects

S Feng

Publications and source records attributed to S Feng.

At least 145 records · Page 8Linked to original sources
PubMed1996

Polymorphism of angiotensin I converting enzyme gene in the older Chinese: linked to ambulatory blood pressure levels and circadian blood pressure rhythm.

This study was undertaken to evaluate the association of polymorphism of angiotensin I converting enzyme (ACE) gene with causal and ambulatory blood pressure in the Chinese population. Genomic DNA was amplified by polymerase chain reaction (PCR) using primers flanking the polymorphic region in intron 16 of the ACE gene. Alleles were detected on agarose gels stained with ethidium bromide. Causal blood pressure was measured in the 133 normotensive and 122 essential hypertensive subjects; of the essential hypertensive subjects, 65 patients underwent non-invasive ambulatory blood pressure. There was no significant difference of ACE genotype and allele frequencies between the hypertensive and normotensive subjects. Non-significant differences were demonstrated in gender, age, causal blood pressure according to the ACE gene genotypes in the 65 patients who underwent non-invasive ambulatory blood pressure; however, effects of the ACE gene genotypes were observed on the derivatives of ambulatory blood pressure, including sleeping blood pressure levels and circadian blood pressure rhythm (p < 0.05). We suggest that insertion/deletion polymorphism of the ACE gene should be associated with blood pressure and ambulatory blood pressure might be superior to causal blood pressure at the molecular level.

Aged↗
PubMed1996

"Coulomb Staircase" at Room Temperature in a Self-Assembled Molecular Nanostructure

Double-ended aryl dithiols [alpha,alpha'-xylyldithiol (XYL) and 4,4'-biphenyldithiol] formed self-assembled monolayers (SAMs) on gold(111) substrates and were used to tether nanometer-sized gold clusters deposited from a cluster beam. An ultrahigh-vacuum scanning tunneling microscope was used to image these nanostructures and to measure their current-voltage characteristics as a function of the separation between the probe tip and the metal cluster. At room temperature, when the tip was positioned over a cluster bonded to the XYL SAM, the current-voltage data showed "Coulomb staircase" behavior. These data are in good agreement with semiclassical predictions for correlated single-electron tunneling and permit estimation of the electrical resistance of a single XYL molecule (approximately18 &plusmn; 12 megohms).

Journal Article↗
PubMed1996

Nuclear localization of a complex of fibroblast growth factor(FGF)-1 and an NH2-terminal fragment of FGF receptor isoforms R4 and R1alpha in human liver cells.

FGF ligands and FGF receptor 1 (FGFR1) appear associated with the nucleus in addition to their extracellular and transmembrane locations. After receptor-dependent internalization in liver cells, radiolabeled 16-kDa FGF-1 appears in a 40-kDa covalent complex with a cellular protein. In this report, we show that in a human hepatoma cell line, HepG2, which expresses both FGFR4 and FGFR1, the 40-kDa complex cross-reacts with antibodies against the ectodomain of both types of receptors. In addition to antibody against FGF-1, a polyclonal antiserum against the three immunoglobulin (Ig)-like loop ectodomain of FGFR4 and a monoclonal antibody to a 19-residue sequence in the NH2-terminus of the NH2-terminal Ig Loop I of the three loop splice variant of FGFR1 (FGFR1alpha) reacts with the complex. A monoclonal antibody against an epitope in FGFR1 downstream of the inter-loop I/II sequence which reacts with intact FGFR1 failed to cross-react with the 40-kDa complex. Cell fractionations and indirect immunofluorescent localization revealed that the 40-kDa complex associates with the particulate fraction of cells, particularly the nucleus and associated cytoskeletal elements. We propose that the NH2-terminal Ig-loop of the three loop isoforms of FGFR, which are generally associated inversely with cell growth, may play a role at or in the nucleus in addition to modification of affinity of the FGFR ectodomain for heparan sulfate and FGF ligand.

Antibodies↗
PubMed1996

Cell-mediated immune response and T-like cells in thymectomized Oncorhynchus mykiss (Walbaum) infected with or vaccinated against the pathogenic haemoflagellate Cryptobia salmositica Katz, 1951.

T-cell-mediated delayed-type hypersensitivity (TDTH) reaction was detected in Cryptobia salmositica-infected intact and thymectomized (2 months post-thymectomy) Oncorhynchus mykiss (Walbaum). Both groups of fish showed significant induration at the site of C. salmositica antigen injection at 8, 12 and 16 weeks post-infection. A significant difference was not observed in TDTH reactions between the thymectomized and intact (control) infected fish. The total numbers of circulating leucocytes detected in infected thymectomized fish were significantly lower than those found in infected shamthymectomized fish. The numbers of T-like cells determined (using alpha-naphthyl acid esterase assay) in thymectomized fish (9 months post-thymectomy) were similar to those seen in intact fish prior to and at 4 weeks after vaccination with an avirulent strain of C. salmositica. At 2 weeks after challenge with the pathogen the numbers of T-like cells in intact vaccinated fish increased significantly (P < 0.01) and remained high for the duration of the study (15 weeks). However, in vaccinated thymectomized fish the numbers of T-like cells remained low after parasite challenge. These results suggest that thymectomy in adult rainbow trout did not lower T-cell-mediated delayed-type hypersensitivity; however, it reduced the numbers of circulating leucocytes and retarded the proliferation of T-like cells after antigenic stimulation.

Animals↗
PubMed1996

A tethered adhesive particle model of two-dimensional elasticity and its application to the erythrocyte membrane.

A new model of two-dimensional elasticity with application to the erythrocyte membrane is proposed. The system consists of a planar array of self-adhesive particles attached to nearest neighbors with flexible tethers. Stretching from the equilibrium dimension is resisted because force is required to dissociate the particle clusters and to decrease the distribution entropy. Release of the external force is accompanied by a contraction as thermal diffusion randomizes the particles and allows interparticle attachments to form again. Analysis of membrane thermodynamics and mechanics under the two-state particle assumption results in a shear softening stress-strain relation. The shear modulus is found proportional to the square root of the surface density of particles, the interparticle adhesive energy, and is inversely proportional to the tether length. Applied to the erythrocyte membrane under the assumption that band 3 tetramer represents the particle and spectrin the tether, the shear modulus predicted corresponds to the measured value when the interparticle adhesive energy is approximately 4.0-5.9 kT, where kT is the Boltzmann constant multiplied by the temperature. This model suggests a mechanism wherein erythrocyte membrane deformability depends on integral protein homomultimeric interactions and can be modulated from the external surface.

Adhesiveness↗
PubMed1996

Molecular basis for the binding of SH3 ligands with non-peptide elements identified by combinatorial synthesis.

BACKGROUND: Protein-structure-based combinatorial chemistry has recently been used to discover several ligands containing non-peptide binding elements to the Src SH3 domain. The encoded library used has the form Cap-M1-M2-M3-PLPPLP, in which the Cap and Mi's are composed of a diverse set of organic monomers. The PLPPLP portion provided a structural bias directing the non-peptide fragment Cap-M1-M2-M3 to the SH3 specificity pocket. Fifteen ligands were selected from > 1.1 million distinct compounds. The structural basis for selection was unknown. RESULTS: The solution structures of the Src SH3 domain complexed with two ligands containing non-peptide elements selected from the library were determined by multidimensional NMR spectroscopy. The non-peptide moieties of the ligands interact with the specificity pocket of Src SH3 domain differently from peptides complexed with SH3 domains. Structural information about the ligands was used to design various homologs, whose affinities for the SH3 domain were measured. The results provide a structural basis for understanding the selection of a few optimal ligands from a large library. CONCLUSIONS: The cycle of protein-structure-based combinatorial chemistry followed by structure determination of the few highest affinity ligands provides a powerful new tool for the field of molecular recognition.

Amino Acid Sequence↗
PubMed1996

P55, an immunogenic but nonprotective 55-kilodalton Borrelia burgdorferi protein in murine Lyme disease.

Immunization of C3H mice with P55 (previously called S1), a 55-kDa Borrelia burgdorferi antigen that is immunogenic after infection, elicited a strong antibody response but did not protect mice against B. burgdorferi challenge. Mice immunized with a P55 fusion protein in complete Freund's adjuvant developed anti-P55 antibodies, detectable at a titer of 1:10,000 by immunoblotting. To determine, if a protective response had been elicited, P55-vaccinated mice were fed upon by ticks infected with B. burgdorferi. The frequency of B. burgdorferi infection was similar in P55-immunized and control mice, and spirochetes were not destroyed within ticks that fed on P55-vaccinated mice. P55 is an immunogenic antigen that does not induce a protective response in the vertebrate or invertebrate host.

Animals↗
PubMed1996

Serum-mediated resolution of Lyme arthritis in mice.

Arthritis and carditis were mildly improved upon adoptive transfer of T cell enriched lymphocyte populations from Borrelia burgdorferi-infected (B. burgdorferi) (immune) compared with naive immunocompetent mice into B. burgdorferi-infected, severe combined immunodeficient (SCID) mice. Despite the relative purity of T cells in transferred cells, recipient mice seroconverted to B. burgdorferi. Thus, the effect could not be attributed to T cells alone. Passive transfer of serum from actively infected immunocompetent mice (immune serum) to SCID mice at the time of or before B. burgdorferi inoculation, or on Days 4, 8, and 12 after inoculation prevented or cured (respectively) infection and disease when examined at 15 days. Transfer of immune serum on Days 12, 16, 20, 24, and 28 did not clear infection at Day 30 but resulted in resolution of arthritis, indicating that immune serum can cause resolution of joint disease. Immune serum treatment could maintain arthritis resolution for up to 60 days. Immune serum from mice infected for 90 days or 15 months both had strong protective, post-infection, and arthritis-modulating activity, whereas hyperimmune serum to heat-killed B. burgdorferi or recombinant outer surface protein (Osp) A protected mice against infection when given on Day 0--but not at later intervals--and did not modulate disease. Immune serum from 90-day infected mice labeled spirochetes in joint tissues of SCID mice by immunohistochemistry, but hyperimmune serum to heat-killed B. burgdorferi or OspA did not. These studies suggest that the biologically active properties of immune serum may be directed toward yet to be defined, in vivo-expressed antigens of B. burgdorferi.

Animals↗
PubMed1996

Multichannel flow electrophoresis and its applications in purification of proteins, enzymes, and antibodies.

Multichannel flow electrophoresis (MFE) is a novel preparative electrophoresis technique designed for continuous separation of proteins based on their different isoelectric points. A computer controlled MFE apparatus is developed and the effects of operation parameters such as electric field strength, buffering pH, and sample introducing flow rate on MFE separation efficiency are investigated. The application of MFE is first exemplified by continuous separation of BSA and HBB mixture. Then continuous purification of urokinase from urine extracts is conducted, in which 10 x 10(4) IU urokinase product whose specific activity is higher than 40,000 IU/mg is obtained per hour. Finally, MFE is applied in the continuous purification of anti-urokinase mouse IgG from mouse serum and yields 2.1 mg mouse IgG per hour with high purity demonstrated by SDS-PAGE. The preliminary results presented have confirmed the workability of MFE and pointed out its high application potential in biochemical process.

Antibodies↗
PubMed1995

Specific interactions outside the proline-rich core of two classes of Src homology 3 ligands.

Two dodecapeptides belonging to distinct classes of Src homology 3 (SH3) ligands and selected from biased phage display libraries were used to investigate interactions between a specificity pocket in the Src SH3 domain and ligant residues flanking the proline-rich core. The solution structures of c-Src SH3 complexed with these peptides were solved by NMR. In addition to proline-rich, polyproline type II helix-forming core, the class I and II ligands each possesses a flanking sequence that occupies a large pocket between the RT and n-Src loops of the SH3 domain. Structural and mutational analyses illustrate how the two classes of SH3 ligands exploit a specificity pocket on the receptor differently to increase binding affinity and specificity.

Adaptor Proteins, Signal Transducing↗
PubMed1995

Lyme disease in human DR4Dw4-transgenic mice.

The evolution of Lyme arthritis in DR4-transgenic mice infected with Borrelia burgdorferi was studied because chronic Lyme arthritis in humans is associated with an increased frequency of the HLA-DR4 allele. B10 nontransgenic and DR4-transgenic mice expressing chimeric human-mouse major histocompatibility complex class II genes in which the human alpha 1 and beta 1 domains of DR4Dw4 replaced the corresponding domains of the mouse I-E(d) were inoculated with B. burgdorferi and examined at up to 180 days for infection and disease. All mice were infected throughout the 180 days, and arthritis evolved to equal severity in transgenic and control mice within 30 days and resolved by day 120. Both groups of mice developed high antibody titers to B. burgdorferi, but antibodies to outer surface proteins A and B were not readily detectable. The DR4Dw4 transgene did not predispose mice to the development of chronic Lyme arthritis.

Alleles↗
PubMed1995

A 55-kilodalton antigen encoded by a gene on a Borrelia burgdorferi 49-kilobase plasmid is recognized by antibodies in sera from patients with Lyme disease.

We have identified a 55-kDa antigen encoded by a gene on a 49-kb plasmid of Borrelia burgdorferi. The screening of a B. burgdorferi DNA expression library (N40 strain) with rabbit anti-B. burgdorferi serum and then with serum from a patient with Lyme disease arthritis revealed a clone that synthesized an antigen that was reactive with both sera. DNA sequence analysis identified an operon with two genes, s1 and s2 (1,254 and 780 nucleotides), that expressed antigens with the predicted molecular masses of 55 and 29 kDa, respectively. Pulsed-field gel electrophoresis showed that the s1-s2 operon was located on the 49-kb plasmid. Recombinant S1 was synthesized as a glutathione S-transferase fusion protein in Escherichia coli. Antibodies to recombinant S1 bound to a 55-kDa protein in lysates of B. burgdorferi, indicating that cultured spirochetes synthesized S1. Thirty-one of 100 Lyme disease patients had immunoglobulin G (IgG) and/or IgM antibodies to S1. IgG antibodies to S1 were detected by enzyme-linked immunosorbent assay and immunoblots in the sera of 21 (21%) of 100 patients with Lyme disease; 11 (27.5%) of the S1-positive samples were from patients (40) with early-stage Lyme disease, and 10 (16.7%) were from patients (60) with late-stage Lyme disease. Fifteen (38.5%) of 40 serum samples from patients with early-stage Lyme disease had IgM antibodies to S1. These data suggest that the S1 antigen encoded by a gene on the 49-kb plasmid is recognized serologically by a subset of patients with early- or late-stage Lyme disease.

Amino Acid Sequence↗
PubMed1995

Structure-function analysis of SH3 domains: SH3 binding specificity altered by single amino acid substitutions.

SH3 domains mediate intracellular protein-protein interactions through the recognition of proline-rich sequence motifs on cellular proteins. Structural analysis of the Src SH3 domain (Src SH3) complexed with proline-rich peptide ligands revealed three binding sites involved in this interaction: two hydrophobic interactions (between aliphatic proline dipeptides in the SH3 ligand and highly conserved aromatic residues on the surface of the SH3 domain), and one salt bridge (between Asp-99 of Src and an Arg three residues upstream of the conserved Pro-X-X-Pro motif in the ligand). We examined the importance of the arginine binding site of SH3 domains by comparing the binding properties of wild-type Src SH3 and Abl SH3 with those of a Src SH3 mutant containing a mutated arginine binding site (D99N) and Abl SH3 mutant constructs engineered to contain an arginine binding site (T98D and T98D/F91Y). We found that the D99N mutation diminished binding to most Src SH3-binding proteins in whole cell extracts; however, there was only a moderate reduction in binding to a small subset of Src SH3-binding proteins (including the Src substrate p68). p68 was shown to contain two Arg-containing Asp-99-dependent binding sites and one Asp-99-independent binding site which lacks an Arg. Moreover, substitution of Asp for Thr-98 in Abl SH3 changed the binding specificity of this domain and conferred the ability to recognize Arg-containing ligands. These results indicate that Asp-99 is important for Src SH3 binding specificity and that Asp-99-dependent binding interactions play a dominant role in Src SH3 recognition of cellular binding proteins, and they suggest the existence of two Src SH3 binding mechanisms, one requiring Asp-99 and the other independent of this residue.

3T3 Cells↗
PubMed1995

[Survival rates in patients with systemic lupus erythematosus].

To assess the impact of demographic and clinical factors on prognosis in patients with systemic lupus erythematosus (SLE), we examined the survival rate by lifetable analysis in 566 patients. All patients were Shanghai citizens who were diagnosed as having SLE in Huashan Hospital between 1959 and 1992. According to American Rheumatism Association's preliminary criteria. The survival rate from the time of SLE onset was 93% at 1 year, 73% at 5 years and 60% at 10 years. On univariate analysis, we found that the following factors worsened the probability of survival; male, neuropsychiatric manifestation, pleurisy-pericarditis, anemia, thrombocytopenia, lymphocytopenia, proteinuria, hematuria, urinary cast, azotemia, decreased endogenous creatinine clearance, increased cholesterol in serum, hypocomplementemia, abnormal electrocardiograph and high corticosteroid dose of treatment. On multivariate analysis, we found the four independent risk factors were male, azotemia, hypocomplementemia and high corticosteroid dose of treatment.

Adolescent↗