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Biomedical subjects

F Y Yang

Publications and source records attributed to F Y Yang.

At least 19 recordsLinked to original sources

Reconstruction control of magnetic properties during epitaxial growth of ferromagnetic Mn3-deltaGa on Wurtzite GaN(0001).

Binary ferromagnetic Mn(3-delta)Ga (1.2<3-delta< or =1.5) crystalline thin films have been epitaxially grown on wurtzite GaN(0001) surfaces using rf N-plasma molecular beam epitaxy. The film structure is face-centered tetragonal with CuAu type-I (L1(0)) ordering with (111) orientation. The in-plane epitaxial relationship to GaN is nearly ideal with [110](MnGa) parallel[1100](GaN) and [112](MnGa) parallel[1120](GaN). We observe magnetic anisotropy along both the in-plane and out-of-plane directions. The magnetic moments are found to depend on the Mn/(Mn+Ga) flux ratio and can be controlled by observation of the surface reconstruction during growth, which varies from 1x1 to 2x2 with increasing Mn stoichiometry.

Journal Article↗

Intermittent exposure to reduced oxygen levels affects prey size selection and consumption in swimming crab Thalamita danae Stimpson.

Portunid crabs Thalamita danae (carapace width: 46-56 mm) were exposed to low oxygen level (4.0 mg O2 l(-1)) and hypoxia (1.5 mg O2 l(-1)) for 6 h each day with three size classes (large: 15.0-19.9 mm, medium: 10.0-14.9 mm, small: 5.0-9.9 mm) of mussels Brachidontes variabilis offered as food. Consumption rate, prey size preference, and prey handling including breaking time, handling time, eating time and prey value, were studied during the time the crabs were exposed to reduced oxygen levels and results were compared with the crabs maintained at high oxygen level (8.0 mg O2 l(-1)) throughout the experiment. Consumption of mussels from all size classes was significantly higher at high oxygen level than at reduced oxygen levels. No mussel size preference was observed for crabs exposed to 4.0 or 8.0 mg O2 l(-1) but those crabs exposed to 1.5 mg O2 l(-1) preferred medium mussels. Both breaking time and handling time increased with mussel size but did not vary with oxygen level. Prey value of each mussel consumed (mg dry wt eaten crab(-1) s(-1)) was calculated by dividing the estimated dry weight of the mussel by the observed handling time. Mean prey value varied significantly with mussel size, with values obtained for large mussels being higher than small mussels at 4.0 and 8.0 mg O2 l(-1); the effect of oxygen level, however, was insignificant. In view of portunid crabs as major predators of mussels, results may help explain dominance of mussels in eutrophic harbours in Hong Kong.

Analysis of Variance↗

Oscillatory exchange bias due to an antiferromagnet with incommensurate spin-density waves.

Oscillatory exchange bias in both magnitude and in sign has been observed in epitaxial (100)Cr/Ni(81)Fe19 bilayers due to the incommensurate spin-density waves in antiferromagnetic (100)Cr layers. Salient effects due to the spin-flip transition between longitudinal and transverse spin-density waves as well as that of expanding wavelength have been observed.

Journal Article↗

Inhibition of thymidylate synthase activity by antisense oligodeoxynucleotide and possible role in thymineless treatment.

Thymidylate synthase (TS) is an important target for chemotherapeutic treatment of cancer. However, efficacy of TS-targeted anticancer drugs is limited by the development of drug resistance as a result of TS gene amplification. In this work, a phosphorothioated antisense oligonucleotide (ODN), designated ATS-2, was used to suppress cellular synthesis of TS. ATS-2 at 0.2 microM concentration was mixed with lipofectin in a charge ratio of 1:1 and was used to treat the human embryonic kidney (HEK) cell line. A reduction of TS mRNA and protein was achieved. Furthermore, a dose-dependent reduction of cumulative viable cells of up to 98% was observed. Flow cytometer analysis of cell cycle progression indicates that ATS-2-treated cells were arrested and went into apoptosis at the S phase, possibly because of thymidine shortage, suggesting that ATS-2 is specifically effective for dividing cells. When used in combination with the anticancer drug FdUrd, ATS-2 exerted a additive inhibitory effect on cellular proliferation. To elucidate the possible role of cellular thymidine kinase (TdR kinase) in ATS-2 treatment, a second cell line, HeLa, was used. Both HEK and HeLa have similar rates of cell division and ODN uptake. In contrast to HEK, which was shown to have very low levels of TdR kinase activity in [(3)H]thymidine incorporation experiments, [(3)H]thymidine incorporation in HeLa was 15-fold greater than that of HEK. We found that HeLa cells were sensitive to FdUrd but were rather resistant to ATS-2. On the contrary, HEK cells were sensitive to ATS-2 but insensitive to FdUrd. Effects of ATS-2 and FdUrd are, therefore, complementary in thymineless treatment too.

Antimetabolites, Antineoplastic↗

Gene synthesis by a LCR-based approach: high-level production of leptin-L54 using synthetic gene in Escherichia coli.

Synthetic genes are very useful in genetic and protein engineering. Here we propose a general method for construction of synthetic genes. Short oligonucleotides are joined through ligase chain reaction (LCR) in high stringency conditions to make "unit fragments" which are then fused to form a full-length gene sequence by polymerase chain reaction. The procedure is simple and accurate and does not place constraints on sequence and length. In this report, a recombinant leptin gene was synthesized according to the codon preference of Escherichia coli. Besides, a substitution of the only Met at position 54 for Leu and an addition of a Met at the N-terminus were introduced in the synthetic gene. The gene was cloned in the pQE-31 expression vector and was expressed in E. coli. A large amount of recombinant leptin containing 6 x His tag was produced and purified by Ni-NTA affinity column. Finally, intact leptin-L54 was released after removing the tag by CNBr cleavage at the Met residue.

Animals↗

Further studies on Zn2+ -mediated domain-domain communication in human erythrocyte band 3.

Human erythrocyte band 3 is purified and reconstituted into vesicles, forming right-side-out proteoliposomes. Zn2+ entrapped inside the proteoliposomes inhibits the anion transport activity of band 3, and removal of the cytoplasmic domain of band 3 is able to diminish Zn2+ inhibition. Thus, the inhibition of activity of band 3 results from the Zn2+ induced conformational change of the cytoplasmic domain, which in turn is transmitted to the membrane domain. The results of intrinsic fluorescence and its quenching by HB and the 35Cl NMR study indicate that the cytoplasmic domain is essential for the conformational change induced by Zn2+. SH-blocking reagents, CH(3)I and GSSG, are used to modify the cytoplasmic domain, where they specifically bind to Cys201 and Cys317. It is observed that the Zn2+ induced inhibition of anion transport activity is blocked. This demonstrates that Cys201 and Cys317 are required in Zn2+ -mediated domain-domain communication.

Anion Exchange Protein 1, Erythrocyte↗

A method for uni-directional reconstitution of human erythrocyte glucose transporter.

The glucose transporter of human erythrocytes (Glut1) was reconstituted into soybean phospholipid liposomes by a method of direct incorporation using the nonionic detergent n-octyl beta-D-glucopyranoside. The reconstituted proteoliposomes were proved to be intact and low ionic permeability. Freeze-fracture electron microscopy study showed that the diameter of the proteoliposomes was about 150 +/- 50 nm and the protein was randomly distributed. The kinetic parameters of the reconstituted transporter were: Km =16.23 mmol/L, Vmax = 34.48 nmol/sec x mg protein. Furthermore, about 90% of the glucose transporter in the reconstituted proteoliposomes were orientated inside-out. Until now it is a more efficient method for uni-directional reconstitution of Glut1 with good reproducibility and higher transport activity.

Detergents↗

The role of ganglioside GM3 in the modulation of conformation and activity of sarcoplasmic reticulum Ca(2+)-ATPase.

Rabbit sarcoplasmic reticulum does contain trace amounts of gangliosides, and the main species is GM3. Incorporation of GM3 into the SR vesicles or addition of it to the soybean phospholipid used for reconstitution of proteoliposomes obviously increased ATP hydrolysis, as well as, Ca2+ uptake activity of sarcoplasmic reticulum Ca(2+)-ATPase. Conformation changes of Ca(2+)-ATPase induced by GM3 were also observed by circular dichroism, intrinsic fluorescence and fluorescence quenching measurements.

Animals↗

Membrane formation and cellular response on the surface of lenses implanted in rabbit eyes.

PURPOSE: To study the pathogenesis of membrane formation and cellular response on the surface of posterior chamber intraocular lenses (IOLs) implanted in rabbits. SETTING: Department of Histology and Embryology, Pathology, Ophthalmology. The First Military Medical University, Guangzhou, P.R. China. METHODS: Thirty rabbits had extracapsular lens extraction and posterior chamber IOL implantation. The IOLs were removed 4, 7, 15, 30, and 90 days postoperatively. Membrane formation and cellular response on IOL surfaces were evaluated using light (n = 25), transmission (n = 5), and scanning electron (n = 5) microscopy. RESULTS: On 30 IOLs, the incidence of cellular adhesion was 100%. Cellular components comprised macrophages, fibroblast-like cells, epithelioid cells, giant cells, ultralarge giant cells, and lymphocytes. A thin, proteinaceous film was also seen on the surface of the IOLs. The membrane of the IOL surface comprised fibrin, collagen fibrils, macrophages, fibroblast-like cells, giant cells, and fibroblasts. CONCLUSION: The findings of this study might apply to humans because cellular elements and membranes have been reported in humans.

Animals↗

Sensitivity of Ca2+ transport of mitochondria to reactive oxygen species.

The relationship between Ca2+ transport and energy transduction of myocardial mitochondria in the presence of reactive oxygen species was investigated. Following treatment with oxygen free radicals [superoxide(O2.-) or hydroxyl radical (.OH)], lipid free radicals in myocardial mitochondrial membrane could be detected by using the method of EPR spin trap. Simultaneously there were obvious alterations in the free Ca2+ ([Ca2+]m) in the mitochondrial matrix; the physical state of membrane lipid; the efficiency of oxidative phosphorylation (ADP/O); the value of the respiratory control ratio (RCR); and the membrane potential of the inner membrane of myocardial mitochondria. If the concentrations of reactive oxygen species were reduced by about 30%, the alterations in the physical state of the membrane lipid and energy transduction of myocardial mitochondria were not observed, but the changes in Ca2+ homeostasis remained. We conclude that Ca2+ transport by myocardial mitochondria is more sensitive to agents such as O2.- or OH, etc. than are oxidation phosphorylation and the respiratory chain.

Animals↗

Lipids may not be involved in the recognition of apocytochrome C during its transportation.

It was shown that apocytochrome c was less insertion into monolayers from outer mitochondrial membrane lipids than into those from microsomal membrane lipids; the alpha-helix content of apocytochrome c induced by small unilamellar vesicles prepared from outer mitochondrial membrane lipids was less than by those from microsomal membrane lipids; the import efficiency of apocytochrome c into large unilamellar vesicles from outer mitochondrial membrane lipids was also lower than into those from microsome membrane lipids. No specific affinity between apocytochrome c and outer mitochondrial membrane lipids could be found. Import of apocytochrome c across the intact mitochondria, sealed outer mitochondrial membrane and microsome membrane vesicles was compared. Results showed that apocytochrome c was accumulated only in mitochondria, but not the other two kinds of vesicles.

Animals↗

V92A mutation altered the folding propensity of chicken apocytochrome c and its interaction with phospholipids.

Chicken apocytochrome c has been shown to possess a much stronger tendency to fold spontaneously in aqueous solution than the equivalent enzyme from other species. In the present work, the amino acid that determines its folding ability was elucidated by site-directed mutagenesis. Wild-type chicken apocytochrome c and three mutants V92A, S103A, and V92A/S103A were expressed in Escherichia coli. The wild-type apoprotein and S103A exhibited the same folding property during dialysis renaturation processes as that chemically prepared from chicken cytochrome c, while those containing V92A mutation did not. Quantitative studies by 2,2,2-trifluoroethanol (TFE) and sodium perchlorate (NaClO4) titration demonstrated that the V92A mutation decreased the helix content that could be induced and confirmed that valine 92 is the major determinant of the folding propensity of chicken apocytochrome c. Furthermore, CD spectra, turbidity measurements, and a translocation assay on a model membrane system showed that the V92A mutation also drastically altered the conformation of apocytochrome c after being incorporated into lipid bilayer and decreased the aggregation of phospholipid vesicles after association of the apoprotein, thus rendering the molecule more competent for translocation across the membrane. Our results showed that a single amino acid substitution could radically alter the folding propensity of an unfolded polypeptide chain and thus influence the conformation following its insertion into phospholipid bilayer.

Animals↗

Effect of ganglioside GM3 on the activity and conformation of reconstituted Ca2+-ATPase.

Trace amounts of gangliosides were found in rabbit skeletal muscle sarcoplasmic reticulum and their main part was shown, by high performance thin layer chromatography. to be GM3. Addition of GM3 to the soybean phospholipids used for reconstitution of proteoliposomes markedly increased ATP hydrolysis as well as Ca2+ uptake activity of sarcoplasmic reticulum Ca2+-ATPase incorporated into the proteoliposomes. Conformation changes of Ca2+-ATPase induced by GM3 were also observed by intrinsic fluorescence and circular dichroism measurements.

Adenosine Triphosphate↗

Transmembrane Ca2+ gradient is essential for high anion transport activity of human erythrocytes.

The role of a transmembrane Ca2+ gradient in anion transport by Band 3 of human resealed erythrocyte ghosts has been studied. The results show that a transmembrane Ca2+ gradient is essential for the conformation of erythrocyte Band 3 with higher anion transport activity. The dissipation of the transmembrane Ca2+ gradient by the ionophore A23187 inhibits the anion transport activity. The extent of this inhibition approaches 90% as the Ca2+ concentration on both sides of the ghost membrane is increased to 1.0 mM and half-maximum inhibitions is observed at 0.25 mM Ca2+. Addition of ATP (0.4 mM) to the resealing medium can partly reestablish the transmembrane Ca2+ gradient by activation of Ca(2+)-ATPase and alleviate the inhibition to some extent. N-ethylmaleimide, an inhibitor of erythrocyte Ca(2+)-ATPase, prevents such restoration. Electron micrographs reveal that numerous larger intramembranous particle can be observed on the P-faces of freeze-fractured resealed ghosts in the absence of a transmembrane Ca2+ gradient.

Anion Exchange Protein 1, Erythrocyte↗

Effect of transmembrane Ca2+ gradient on the coupling of beta-adrenergic receptors and adenylyl cyclase.

In order to investigate the effect of transmembrane Ca2+ gradient on Gs mediated coupling of beta-AR and adenylyl cyclase, beta-AR from duck erythrocytes and Gs and adenylyl cyclase from bovine brain cortices were co-reconstituted into asolectin liposomes with different transmembrane Ca2+ gradient. These proteoliposomes were proven to be impermeable to water-soluble substances. The results obtained indicate that a physiological transmembrane Ca2+ gradient (1000-fold) is essential for higher stimulation of adenylyl cyclase by hormone-activated beta-AR via coupling to Gs and can be further enhanced by the decrease of such Ca2+ gradient within certain range (100 fold) following Ca2+ influx into cells during signal transduction. Fluorescence polarization of DPH revealed that transmembrane Ca2+ gradient modulates adenylyl cyclase and its stimulation by hormones through mediating a change in lipid fluidity. Correspondent conformational changes of beta-AR were also detected from the fluorescence spectra and quenching of Acrylodan-labelled beta-AR in those proteoliposomes. It is suggested that a proper transmembrane Ca2+ gradient is essential for the optimal fluidity of the phospholipid bilayer in the proteoliposomes, which favors the formation of a suitable conformation of the reconstituted beta-AR and thus promotes the stimulation of adenylyl cyclase activities by hormone-activated beta-AR via Gs.

Adenylyl Cyclases↗