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Biomedical subjects

S Y Mao

Publications and source records attributed to S Y Mao.

At least 19 recordsLinked to original sources

alpha-Adducin gene and essential hypertension in China.

Adducin is a membrane skeletal protein that is involved in the regulation of membrane ion transport and cellular signal transduction. Essential hypertension has been linked to alpha-adducin gene locus, and association of a polymorphism of the gene has been found in some studies, but results of linkage or association studies on alpha-adducin gene are controversial among different populations. This study was designed to examine the linkage between alpha-adducin gene locus and essential hypertension and to reveal the relationship between an alpha-adducin gene polymorphism (Gly460Trp) and essential hypertension in a Chinese population. For the linkage study, one hundred and six Chinese nuclear families were recruited, including 417 hypertensive patients in all 474 individuals. Those samples were genotyped at D4S412 and D4S3038. The distances between the two microsatellite markers and the alpha-adducin gene locus are less than 3cM. Parametric, non-parametric linkage (NPL) analyses using the GENEHUNTER software were carried out. Sib transmission-dise- quilibrium test (S-TDT), as well as transmission-disequilibrium test (TDT). was also implemented with TDT/S-TDT Program 1.1. Serum levels of uric acid, creatinine, blood urea nitrogen (BUN), fasting glucose and lipids were determined as phenotypes. In an association study, 138 hypertensive and 121 normotensive subjects were genotyped at Gly460Trp of the alpha-adducin gene to examine a possible association between this polymorphism and blood pressure or other phenotypes. We fail to find the linkage between the two markers and essential hypertension by parametric, NPL analysis or TDT/S-TDT study. With the use of the simple association and the multivariate logistic regression analyses, we also fail to reveal a significant association between the Gly460Trp polymorphism in alpha-adducin gene and the blood pressure variation, or blood biochemical indices studied. The frequency of the 460Trp allele in Chinese (46-48%) is similar to that found in Japanese (54-60%) while the allele frequency is less common in Caucasian (13%-23%). These findings suggest that in our Chinese population, alpha-adducin 460Trp variant may not play an important role in the etiology of EH. And the negative results of linkage and TDT/ S-TDT further supports this conclusion.

Adult↗

Selective deletion of antigen-specific, activated T cells by a humanized MAB to CD2 (MEDI-507) is mediated by NK cells.

CD2 is a 50-kDa transmembrane glycoprotein that plays an important role in T and natural killer (NT) lymphocyte functions. CD2 serves as both an adhesion molecule and as a costimulatory molecule through interactions with its ligand, CD58, on antigen presenting or target cells. Consistent with earlier studies using a rat anti-CD2 mAb, we have shown that treatment of alloantigen stimulated T lymphocytes with a humanized mAb, MEDI-507 (IgG1, kappa), induced hyporesponsiveness to subsequent stimulation with alloantigen but not to mitogen (phytohemagglutinin). Fluorescence-activated cell sorting analysis of cells from mixed lymphocyte reaction (MLR) treated with MEDI-507 revealed pronounced deletion of T and NK cells, consistent with lack of proliferation in the MLR. MEDI-507 F(ab')2 fragments did not have inhibitory activity or induce deletion of lymphocytes in the MLR. Removal of the NK cell subset by magnetic bead depletion using anti-CD16 and anti-CD56 mAbs eliminated both the T cell deletion and the inhibitory effect. Reconstitution of NK depleted responder populations using autologous NK cells restored the MEDI-507-mediated deletion activity to levels measured in the original MLR. Formaldehyde-fixed NK cells failed to mediate the MEDI-507-induced deletion effect. Altogether, our studies indicate that activated T cells with MEDI-507 bound to CD2 are preferential targets for autologous NK cells through a nonapoptotic cytotoxic mechanism.

Adult↗

Assessment of radionuclide contents in food in Hong Kong.

Baseline values of concentrations of the natural radionuclides (238U, 226Ra, 228Ra/232Th, 210Pb) and artificial radionuclides (137Cs, 60Co) in food and drinks (tap water, milk, and water-based drinks) were determined by gamma spectroscopy. All food and drinks were found to contain detectable 40K contents: 0.1 to 160 Bq kg(-1) (fresh mass) for food and 0.006 to 61 Bq L(-1) for drinks. Most of the other natural radionuclides in solid food were found to have contents below the minimum detectable activities (MDA). More samples in the leafy vegetable, tomato, carrot and potato categories contained detectable amounts of 228Ra than the meat, cereal, and fish categories, with concentrations up to 1.2 Bq kg(-1) for the former categories and 0.35 Bq kg(-1) for the latter categories. The 238U and 226Ra radionuclides were detectable in most of the water-based drink samples (reaching 0.22 and 0.015 Bq kg(-1), respectively), and the 228Ra and 210Pb radionuclides were detectable in fewer water-based drink samples. The 137Cs contents in solid food were detectable in most of the solid food samples (reaching 0.59 Bq kg(-1)), but in drinks the 137Cs contents were very low (the maximum value is 0.23 Bq L(-1)) and normally lower than the MDA values. Nearly all the 60Co contents in food and drinks were below the MDA values and their contents were below those of 137Cs. The results indicate that drinks provide higher intake of natural radionuclides than solid food, but the solid food provides higher intake of 137Cs than drinks. From the measured concentrations, estimates were made on the consumption rate of radionuclides from food and drinks. The health effects due to the ingestion of the radionuclides were evaluated by calculating the committed effective dose. The committed effective dose due to the ingestion of natural radionuclides was estimated to be 69 microSv and 280 microSv for an adult male and female, respectively. The committed effective dose due to the ingestion of 137Cs and 60Co was estimated to be 0.5 microSv for both an adult male and female. The contributions from these two artificial radionuclides were less than those from the natural radionuclides by a factor of more than 100 and were also below the 1 mSv limit set in the new ICRP recommendations.

Adult↗

Assessment of the transfer of 137Cs in three types of vegetables consumed in Hong Kong.

A dynamic food chain model has been built for the modeling of the transfer of 137Cs in three types of vegetables consumed in Hong Kong, namely, white flowering cabbage (Brassica chinensis), head lettuce (Lactuca sativa) and celery (Apium graveolens). Some parameters have been estimated from the experimental data obtained in this work. The experimental data include the transfer factors of 137Cs from soil to the different vegetable species which are determined through high resolution gamma spectrometry, maximum crop biomasses for the vegetable species, the dry-to-fresh ratios for the vegetable species, the bulk density of soil layers and the average concentration of 137Cs in air. The derived parameters include the deposition rate and the root uptake rate, information for tillage, the logistic growth model and radionuclide concentrations in vegetables. The dynamic food chain model is solved by the Birchall-James algorithm to give the 137Cs concentration in subsurface soil, from the 0.1-25 cm soil layer, and the 137Cs concentration in harvested and unwashed vegetables. As validation of the model and parameters, the concentrations obtained experimentally and from the model are compared and are found to be in good agreement.

Apiaceae↗

Characterization of protein-tyrosine phosphatases that dephosphorylate the high affinity IgE receptor.

An early event that follows aggregation of the high affinity receptor for IgE (FcepsilonRI) is the phosphorylation of protein tyrosines, especially those on the beta- and gamma-subunits of the receptor. Disaggregation of the receptors leads to their rapid dephosphorylation, but even stably aggregated receptors undergo continual rounds of phosphorylation and dephosphorylation. We developed assays to study dephosphorylation of the receptors and other cellular proteins. Whole cell extracts dephosphorylated both subunits of the receptors rapidly and were as active against aggregated as against disaggregated FcepsilonRI. Upon disaggregation, the in vivo dephosphorylation of the FcepsilonRI and several other proteins followed first-order kinetics with closely similar rate constants despite substantial differences in the extent of phosphorylation. These results suggest that the level of phosphorylation of FcepsilonRI is largely controlled by the aggregation-induced action of kinase(s) and not from changes in susceptibility to or activity of the phosphatases. Much of the total phosphatase is lost when the cells are permeabilized, but the rate of dephosphorylation of disaggregated FcepsilonRI was comparable in intact and permeabilized cells. Thus, much of the activity utilized by the cell to dephosphorylate the FcepsilonRI is likely to be associated with the plasma membrane.

Animals↗

A study of radioactivities in six types of fish consumed in Hong Kong.

The contents of natural and artificial radionuclides in two types of fresh water fish and four types of marine fish commonly consumed by the Hong Kong population have been measured using HPGe gamma-ray spectrometry. The two types of fresh water fish are grey mullet (mugil cephalus) and grass carp (ctenopharyngodon idellus). The four types of marine fish are white pomfret (pampus argenteus), red bullseye (priacanthus macracanthus), golden thread (nemipterus) and ginkgo (gymnocranius griseus). All measurements of U and Th chain radionuclides in the samples have yielded values below MDA except for positive detection of 228Ra (assuming 228Ra to be in equilibrium with 228Ac) in grey mullet and bullseye perch. The 40K and 137Cs radionuclides were detectable in all samples, with values of 0.01-0.2 and 41.23-111.47 Bq/kg fresh respectively. The different levels of radioactivity in different species of fish is suggested to be due to differences in metabolism and feeding patterns. The radionuclide 60Co, the only other artificial radionuclide to be investigated herein, recorded values below MDA for most of the samples; observable values ranged from 0.01 to 0.04 Bq/kg fresh. Committed effective doses due to ingestion of radionuclides from fish have been estimated to be 1.2 (male) and 6.4 (female) microSv/yr for natural radionuclides; 0.027 (male) and 0.026 (female) microSv/yr for artificial radionuclides.

Animals↗

Kinetics of tyrosine phosphorylation when IgE dimers bind to FC epsilon receptors on rat basophilic leukemia cells.

Previously, we demonstrated that aggregates of the high affinity receptor for IgE (Fc epsilon RI), formed by the binding of chemically cross-linked oligomers of IgE, continue to signal early and late cellular responses long after the formation of new aggregates is blocked. In the present work, we explore quantitatively the relationship between aggregation of the receptors and one of the earliest biochemical changes this initiates. We compare the time course of aggregate formation, inferred from studies of the binding of dimers of IgE, and the time course of phosphorylation of tyrosines on receptor subunits when the receptors are aggregated. A simple model does not fit the data. It appears that aggregates formed late in the response are less effective signaling units than those formed initially. We propose new explanations for the persistence of the response and the unusual kinetics.

Animals↗

Effects of subunit mutation on the rotational dynamics of Fc epsilon RI, the high affinity receptor for IgE, in transfected cells.

Erythrosin-labeled immunoglobulin E (IgE) and time-resolved phosphorescence anisotropy were used to monitor the rotational dynamics of transfected wild-type (alpha beta gamma 2) and four mutant Fc epsilon RI receptors in the monomeric and dimeric state on P815 cells. Erythrosin-IgE bound to Fc epsilon RI on cells transfected with either beta or gamma subunits with truncated COOH-terminal cytoplasmic segments exhibit faster rotational motion than when bound to Fc epsilon RI on cells transfected with wild-type subunits. Deletion of the NH2-terminal cytoplasmic segment of the beta subunit or the COOH-terminal cytoplasmic segment of the alpha subunit does not cause any significant change in the anisotropy decay. Dimers of IgE-receptor complexes formed with anti-IgE monoclonal antibody B1E3 exhibit substantially slower anisotropy decays for all the receptor constructs used, including a receptor construct that only contains the ectodomain of the alpha subunit anchored to Chinese hamster ovary (CHO) cell membranes through a lipid tail. This loss of rotational motion of dimeric IgE-Fc epsilon RI complexes may be due to nonspecific entanglement or to specific interactions involving IgE or the extracellular portion of alpha. The results suggest that the beta and gamma subunits of the tetrameric alpha beta gamma 2 receptor participate in interactions with other membrane components even in the absence of receptor aggregation. The loss of such interactions may be related to the functional impairments previously determined for these mutants.

Animals↗

Chemical cross-linking of IgE-receptor complexes in RBL-2H3 cells.

Aggregation of the high-affinity receptors for IgE (Fc epsilon RI) on mast cells activates nonreceptor kinases and other enzymes, thereby initiating a complex biochemical cascade. We have employed a chemical cross-linker in order to stabilize the association of Fc epsilon RI with other cellular proteins that are involved in the early events. We reacted the water-soluble, membrane-impermeant chemical cross-linker 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP) with permeabilized rat mucosal mast cells of the RBL line and analyzed immunoprecipitates of the receptor in detergent lysates of cells that had biosynthetically incorporated [35S]cysteine. Gel electrophoresis revealed substantial amounts of nonreceptor components only when the cells had been reacted with DTSSP. Receptors isolated from cells whose receptor-bound IgE had been aggregated with antigen prior to cross-linking yielded a similar pattern of 35S-labeled proteins. However, when the cells had also been exposed to [gamma-32P]ATP, the proteins associated with the cross-linked, aggregated receptors revealed enhanced incorporation of 32P compared to those associated with cross-linked, unaggregated receptors. A variety of antibodies were used to try to identify the associated proteins. Of those tested for, two, the src-like kinase p53/56lyn and the delta isoform of protein kinase C, were associated with the cross-linked Fc epsilon RI in amounts much greater than could be accounted for by nonspecific cross-linking.

Animals↗

Aggregation of the high-affinity IgE receptor and enhanced activity of p53/56lyn protein-tyrosine kinase.

Aggregation of the receptor with high affinity for IgE (Fc epsilon RI) on the surface of mast cells and basophils stimulates phosphorylation of protein tyrosines, a process in which p53/56lyn kinase has been implicated. We measured the association between Fc epsilon RI and the kinase, using chemical crosslinking to stabilize their interaction. In the rat basophilic leukemia mast cell line, 3-4%, and at most 20%, of Fc epsilon RI appear to be associated with the kinase prior to aggregation, even though there is an excess of total cell lyn kinase. Aggregating the Fc epsilon RI causes three to four times more of the kinase to associate with receptors, a process requiring a prior phosphorylation step. In an in vitro assay, the lyn associated with the aggregated receptors becomes disproportionately more phosphorylated than would be predicted from the amount of lyn associated with the receptors. These and other data are consistent with a model in which aggregation of the receptor leads to its transphosphorylation by constitutively associated lyn kinase. We propose that additional molecules of this kinase are thereby recruited and that this markedly enhances transphosphorylation of tyrosine on the receptor and associated proteins, thereby initiating a cascade of further biochemical changes. This model is also consistent with data on receptors such as the clonotypic receptors on B and T lymphocytes, which share structural and functional features with Fc epsilon RI.

Animals↗

Dynamics of signal transduction after aggregation of cell-surface receptors: studies on the type I receptor for IgE.

Many ligands stimulate cellular responses by aggregating the cell-surface receptors to which they are bound. We investigated several mechanistic questions related to aggregation of receptors by using the high-affinity receptor for IgE (Fc epsilon RI) on mast cells as a model system. We briefly exposed cells to covalently cross-linked oligomers of IgE and then added excess monomeric IgE to prevent further aggregation. Early events were examined by monitoring the phosphorylation of protein tyrosines; later events were examined by monitoring secretion. We found that aggregated receptors continue to signal both late and early events in the absence of formation of new aggregates. Additional experiments suggested that the clustered receptors undergo a dynamic process of phosphorylation and dephosphorylation. Our findings suggest that for these and related receptors that function by aggregation, the persistence of signal transduction is directly related to the intrinsic affinity of the ligand for the individual receptor.

Animals↗