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Biomedical subjects

W Yi

Publications and source records attributed to W Yi.

At least 19 recordsLinked to original sources

Detecting the breached-pair phase in a polarized ultracold Fermi gas.

We propose a method for the experimental detection of a new quantum phase, the breached-pair state, in a strongly interacting ultracold Fermi gas with population imbalance. We show that through the time-of-flight Raman imaging, the presence of such a phase can be unambiguously determined with a measurement of the momentum-space phase separation of the minority spin component. To guide the experimental efforts, the momentum-space density profiles are calculated under typical experimental conditions.

Journal Article↗

Biological denitrification in a sequencing batch reactor.

Biological denitrification was studied in an SBR to investigate the performance. The experimental results showed that granular sludge with high activity and good settling ability can be developed smoothly in thereactor. With the granular sludge, high nitrate loading rate (0.48 kg NO3-N dm(-3) d(-1)) and removal efficiency (more than 99%) can be achieved. Scanning electronic microscope examination showed that only rod-shape bacteria existed on and within the granules, which confirmed indirectly that filamentous bacteria do not utilize nitrate and nitrite as electron acceptors. SBR is an alternative and effective technology for biological denitrification.

Ammonia↗

Tunneling into multiwalled carbon nanotubes: coulomb blockade and the Fano resonance.

Tunneling spectroscopy measurements of single tunnel junctions formed between multiwalled carbon nanotubes (MWNTs) and a normal metal are reported. Intrinsic Coulomb interactions in the MWNTs give rise to a strong zero-bias suppression of a tunneling density of states that can be fitted numerically to the environmental quantum-fluctuation theory. An asymmetric conductance anomaly near zero bias is found at low temperatures and interpreted as Fano resonance in the strong tunneling regime.

Journal Article↗

Initial investigations of 99mTc-labeled morpholinos for radiopharmaceutical applications.

This laboratory is evaluating phosphorothioate deoxyribonucleic acids (DNAs) and peptide nucleic acids (PNAs) for a variety of nuclear medicine applications. Morpholinos (MORFs) are a new class of oligomers with a nuclease-resistant, nonionic and water-soluble phosphorodiamidate backbone. We now report on the in vitro and in vivo properties of MORFs labeled with technetium-99m. Both 15-mer and 18-mer MORFs were obtained, each with a primary amine attached to the 3' equivalent end via a three-carbon beta-alanine linker. The amine was used to conjugate with NHS-MAG3 for 99mTc radiolabeling. By surface plasmon resonance at room temperature, the association rate constant for hybridization of the 18-mer MORF to its complementary oligomer (cMORF) was equivalent to that of DNAs and PNAs of comparable length. Hybridization of 99mTc-MORF in vitro to free cMORF, to a cMORF polymer and to cMORF beads was nearly quantitative under a variety of conditions. Kinetic studies in vitro at room temperature showed rapid (2-5 min) and nearly quantitative (90%) binding to cMORF beads. Using size-exclusion high-performance liquid chromatography, the stability of the 99mTc-MORF was found to be greater than 85% over 24 h in 37 degrees C serum with minimal protein binding. In normal mice, the 99mTc-MORF showed rapid pharmacokinetics, with only 21% and 8% remaining in the whole body at 3 and 24 h post administration, respectively. In vivo targeting with 99mTc-MORF of cMORF beads in one thigh of normal mice compared to control beads in the other thigh showed target/control thigh ratios of 2-10 between 3 and 24 h. These results demonstrate that MORF oligomers are capable of in vivo hybridization. Their properties of hybridization affinity and kinetics and their in vivo stability and pharmacokinetics make them suitable subjects for in vivo studies.

Animals↗

New mutations in the neuronal ceroid lipofuscinosis genes.

Thirty-eight mutations and seven polymorphisms have recently been reported in the genes underlying the neuronal ceroid lipofuscinoses (NCLs) including 11 new mutations described here. A total of 114 mutations and 28 polymorphisms have now been described in the five human genes identified which cause NCL. Thirty-eight mutations are recorded for CLN1/PPT; 40 for CLN2/TTP-1, 31 for CLN3, four for CLN5, one for CLN8. Two mutations have been described in animal genes (cln8/mnd, CTSD). All mutations in NCL genes are contained in the NCL Mutation Database (http://www.ucl.ac.uk/NCL).

Child↗

[A preliminary study on the organic carbon weathering fluxes in Beijiang River drainage].

Riverine water samples were collected from Hekou hydrometric station of Beijiang River in five hydrological seasons. The samples were analyzed of their organic carbon and total suspended substance, which demonstrated that the concentrations of particulate organic carbon (POC) changed synchronically with the concentrations of total suspended substance (TSS) in riverine water. The correlation between the concentrations of TSS and dissolved organic carbon (DOC), however, is sometimes positive and sometimes negative, and the correlation is not so notable as that between TSS and POC. With the concentration of total suspended substances increasing, the quality partition of organic carbon in total suspended substance decreased in a logarithm tendency. The transportation of organic carbon, especially of DOC, mainly takes place in flood peak periods. The weathering flux of organic carbon in Beijiang River drainage basin is about 10.01 x 10(6) g.km-2.a-1, which is constituted by 6.54 x 10(6) g.km-2.a-1 of POC and 3.47 x 10(6) g.km-2.a-1 of DOC. This pattern of organic carbon weathering flux in Beijiang drainage basin is consistent with that of most of the monsoon drainage basins.

Carbon↗

LMP1 activates NF-kappa B via degradation of I kappa B alpha in nasopharyngeal carcinoma cells.

OBJECTIVE: To elucidate the mechanisms by which Epstein-Barr virus-encoded latent membrane protein 1 activates NF-kappa B in nasopharyngeal carcinoma cells. METHODS: A tetracycline-regulated LMP1-expressing nasopharyngeal carcinoma cell line, Tet-on-LMP1-HNE2, was used as the cell model. The kinetics of the expression of proteins, including LMP1, I kappa B alpha and I kappa B beta, was analyzed by Western blotting. The subcellular localization of NF-kappa B (p65) was detected by indirect immunofluorescence assay. The NF-kappa B transactivity was studied by transient transfection and reporter gene assay. RESULTS: I kappa B alpha was phosphorylated and degraded after the inducible expression of LMP1, although the total protein levels remained stable. The steady-state level of total I kappa B beta protein may have resulted from the initiation of an autoregulation loop after the activation of NF-kappa B. No change in the I kappa B beta level was detected. NF-kappa B (p65) was translocated from the cytoplasm to the nucleus following degradation of I kappa B alpha. After the introduction of the dominant-negative mutant of I kappa B alpha (Del 71) into Tet-on-LMP1-HNE2 cells, both nuclear translocation and transactivation of NF-kappa B induced by LMP1 was significantly inhibited. CONCLUSIONS: The results indicated that in nasopharyngeal carcinoma cells, LMP1 activated NF-kappa B via phosphorylation and degradation of I kappa B alpha, but not I kappa B beta. The dominant-negative mutant of I kappa B alpha (Del 71) could completely inhibit both the nuclear translocation and transactivation of NF-kappa B induced by LMP1.

Biological Transport↗

[Epstein-Barr virus latent membrane protein 1 induces the telomerase activity in human nasopharyngeal epithelial cells].

Telomerase activation has been linked to cell immortalization in vitro and tumorigenicity in vivo. In this study, for the first, we reported that Epstein-Barr virus activated the telomerase activity of human nasopharyngeal epithelial cells in the early stage of immortalization as tested by the PCR-ELISA. The telomerase activity in nasopharyngeal epithelial cells was only observed in presenescent cells. It was implicated that Epstein-Barr virus induced the escape of nasopharyngeal epithelial cells from senescence via the activation of telomerase. We further showed that telomerase activation in infected cells was dependent on the protein level of latent membrane protein 1 (LMP1) encoded by Epstein-Barr virus using a Tetracycline regulatory cell line expressing LMP1, pTet-on-LMP1-HNE2. The activity of telomerase in nasopharyngeal cells was decreased when the protein level of LMP1 was blocked by antisense LMP1 plasmid DNA. And the activity of telmerase was also related to the carboxyl terminus of LMP1. It was implicated that the ability of Epstein-Barr virus to suppress senescence is associated with telomerase activation by LMP1.

Cell Transformation, Viral↗

Mab-3 is a direct tra-1 target gene regulating diverse aspects of C. elegans male sexual development and behavior.

Sex determination is controlled by global regulatory genes, such as tra-1 in Caenorhabditis elegans, Sex lethal in Drosophila, or Sry in mammals. How these genes coordinate sexual differentiation throughout the body is a key unanswered question. tra-1 encodes a zinc finger transcription factor, TRA-1A, that regulates, directly or indirectly, all genes required for sexual development. mab-3 (male abnormal 3), acts downstream of tra-1 and is known to be required for sexual differentiation of at least two tissues. mab-3 directly regulates yolk protein transcription in the intestine and specifies male sense organ differentiation in the nervous system. It encodes a transcription factor related to the products of the Drosophila sexual regulator doublesex (dsx), which also regulates yolk protein transcription and male sense-organ differentiation. The similarities between mab-3 and dsx led us to suggest that some aspects of sex determination may be evolutionarily conserved. Here we find that mab-3 is also required for expression of male-specific genes in sensory neurons of the head and tail and for male interaction with hermaphrodites. These roles in male development and behavior suggest further functional similarity to dsx. In male sensory ray differentiation we find that MAB-3 acts synergistically with LIN-32, a neurogenic bHLH transcription factor. Expression of LIN-32 is spatially restricted by the combined action of the Hox gene mab-5 and the hairy homolog lin-22, while MAB-3 is expressed throughout the lateral hypodermis. Finally, we find that mab-3 transcription is directly regulated in the intestine by TRA-1A, providing a molecular link between the global regulatory pathway and terminal sexual differentiation.

Animals↗

Structure of the human palmitoyl-protein thioesterase-2 gene (PPT2) in the major histocompatibility complex on chromosome 6p21.3.

Palmitoyl-protein thioesterase-2 (PPT2) is a homolog of PPT1, the enzyme that is deficient in the lysosomal storage disorder, infantile neuronal ceroid lipofuscinosis (NCL). As a first step toward determining whether mutations in the gene encoding PPT2 (PPT2) are associated with any of the molecularly uncharacterized forms of NCL, we report here the structure and chromosomal localization of human PPT2. PPT2 spans about 10 kb and is composed of nine exons. One major (2.0 kb) and two minor (7.0 and 2.8 kb) mRNAs are transcribed from the gene, and the larger transcripts appear to be messenger RNAs in which PPT2 exons are spliced into a downstream gene encoding a homolog of human latent transforming growth factor-beta binding protein (human LTBP). PPT2 is located in the human major histocompatibility class III locus on chromosome 6p21.3, a position that rules out PPT2 as the causative gene in any of the NCLs at defined chromosomal loci. No mutations were detected by SSCP analysis in a preliminary analysis of 12 subjects referred with a suspected diagnosis of infantile NCL who had normal PPT activity. However, five single nucleotide polymorphisms were found in unrelated normal individuals. These polymorphisms (and a microsatellite discovered within PPT2) will aid in the further delineation of the possible role of PPT2 in lysosomal storage disorders of unknown etiology.

Amino Acid Sequence↗

Genotype-phenotype correlations in neuronal ceroid lipofuscinosis due to palmitoyl-protein thioesterase deficiency.

The infantile form of neuronal ceroid lipofuscinosis (NCL) has been well studied in Finland, where there is a high carrier frequency (1:70) for a single mutation in the causative gene, CLN1, or PPT. We have recently studied a group of 29 NCL subjects in the United States with palmitoyl-protein thioesterase (PPT) deficiency and described 19 different CLN1/PPT mutations in our population. In this report, we present a review of our previous findings, including a more detailed analysis of phenotype-genotype correlations, and present previously unpublished data concerning the clinical manifestations of the disorder in children of families with multiple affected members. Our studies indicate that about half of PPT-deficient patients in the United States are very similar to Finnish infants with INCL, but that a different mutation (R151X) accounts for 40% of U.S. alleles. The Finnish mutation (R122W) is rare in the United States. The other half of U.S. PPT-deficient patients develop symptoms after the age of 2 years, much later than Finnish patients. One common mutation (the "Scottish" allele, T75P) accounts for 13% of alleles and results in a juvenile-onset phenotype that is clinically indistinguishable from JNCL with CLN3 mutations. Other rare mutations were also associated with JNCL phenotypes, such as D79G and G250V. A preliminary expression study of two of these mutant enzymes supports the conclusion that juvenile-onset NCL (JNCL with GROD) is caused by missense mutations in the PPT gene that result in mutated enzymes with residual PPT enzyme activity.

Age of Onset↗

Similarity of DNA binding and transcriptional regulation by Caenorhabditis elegans MAB-3 and Drosophila melanogaster DSX suggests conservation of sex determining mechanisms.

Although most animals occur in two sexes, the molecular pathways they employ to control sexual development vary considerably. The only known molecular similarity between phyla in sex determination is between two genes, mab-3 from C. elegans, and doublesex (dsx) from Drosophila. Both genes contain a DNA binding motif called a DM domain and they regulate similar aspects of sexual development, including yolk protein synthesis and peripheral nervous system differentiation. Here we show that MAB-3, like the DSX proteins, is a direct regulator of yolk protein gene transcription. We show that despite containing different numbers of DM domains MAB-3 and DSX bind to similar DNA sequences. mab-3 mutations deregulate vitellogenin synthesis at the level of transcription, resulting in expression in both sexes, and the vitellogenin genes have potential MAB-3 binding sites upstream of their transcriptional start sites. MAB-3 binds to a site in the vit-2 promoter in vitro, and this site is required in vivo to prevent transcription of a vit-2 reporter construct in males, suggesting that MAB-3 is a direct repressor of vitellogenin transcription. This is the first direct link between the sex determination regulatory pathway and sex-specific structural genes in C. elegans, and it suggests that nematodes and insects use at least some of the same mechanisms to control sexual development.

Amino Acid Sequence↗

Molecular genetics of palmitoyl-protein thioesterase deficiency in the U.S.

Mutations in a newly described lysosomal enzyme, palmitoyl-protein thioesterase (PPT), were recently shown to be responsible for an autosomal recessive neurological disorder prevalent in Finland, infantile neuronal ceroid lipofuscinosis. The disease results in blindness, motor and cognitive deterioration, and seizures. Characteristic inclusion bodies (granular osmiophilic deposits [GROD]) are found in the brain and other tissues. The vast majority of Finnish cases are homozygous for a missense mutation (R122W) that severely affects PPT enzyme activity, and the clinical course in Finnish children is uniformly rapidly progressive and fatal. To define the clinical, biochemical, and molecular genetic characteristics of subjects with PPT deficiency in a broader population, we collected blood samples from U.S. and Canadian subjects representing 32 unrelated families with neuronal ceroid lipofuscinosis who had GROD documented morphologically. We measured PPT activity and screened the coding region of the PPT gene for mutations. In 29 of the families, PPT deficiency was found to be responsible for the neurodegenerative disorder, and mutations were identified in 57 out of 58 PPT alleles. One nonsense mutation (R151X) accounted for 40% of the alleles and was associated with severe disease in the homozygous state. A second mutation (T75P) accounted for 13% of the alleles and was associated with a late onset and protracted clinical course. A total of 19 different mutations were found, resulting in a broader spectrum of clinical presentations than previously seen in the Finnish population. Symptoms first appeared at ages ranging from 3 mo to 9 yr, and about half of the subjects have survived into the second or even third decades of life.

Adolescent↗

Involvement of the Src homology 2-containing tyrosine phosphatase SHP-2 in growth hormone signaling.

Growth hormone (GH) signaling requires activation of the GH receptor (GHR)-associated tyrosine kinase, JAK2. JAK2 activation by GH is believed to facilitate initiation of various pathways including the Ras, mitogen-activated protein kinase, STAT, insulin receptor substrate (IRS), and phosphatidylinositol 3-kinase systems. In the present study, we explore the biochemical and functional involvement of the Src homology 2 (SH2)-containing protein-tyrosine phosphatase, SHP-2, in GH signaling. GH stimulation of murine NIH 3T3-F442A fibroblasts, cells that homologously express GHRs, resulted in tyrosine phosphorylation of SHP-2. As assessed specifically by anti-SHP-2 coimmunoprecipitation and by affinity precipitation with a glutathione S-transferase fusion protein incorporating the SH2 domains of SHP-2, GH induced formation of a complex of tyrosine phosphoproteins including SHP-2, GHR, JAK2, and a glycoprotein with properties consistent with being a SIRP-alpha-like molecule. A reciprocal binding assay using IM-9 cells as a source of SHP-1 and SHP-2 revealed specific association of SHP-2 (but not SHP-1) with a glutathione S-transferase fusion incorporating GHR cytoplasmic domain residues 485-620, but only if the fusion was first rendered tyrosine-phosphorylated. GH-dependent tyrosine phosphorylation of SHP-2 was also observed in murine 32D cells (which lack IRS-1 and -2) stably transfected with the GHR. Further, GH-dependent anti-SHP-2 coimmunoprecipitation of the Grb2 adapter protein was detected in both 3T3-F442A and 32D-rGHR cells, indicating that biochemical involvement of SHP-2 in GH signaling may not require IRS-1 or -2. Finally, GH-induced transactivation of a c-Fos enhancer-driven luciferase reporter in GHR- and JAK2-transfected COS-7 cells was significantly reduced when a catalytically inactive SHP-2 mutant (but not wild-type SHP-2) was coexpressed; in contrast, expression of a catalytically inactive SHP-1 mutant allowed modestly enhanced GH-induced transactivation of the reporter in comparison with that found with expression of wild-type SHP-1. Collectively, these biochemical and functional data imply a positive role for SHP-2 in GH signaling.

3T3 Cells↗

Uptake of moderately hydrophobic chlorophenols from water by semipermeable membrane devices (SPMDs) and by goldfish (Crassius auratus).

Semipermeable membrane devices (SPMDs) filled with triolein and goldfish (Crassius auratus) were simultaneously exposed to nine moderately hydrophobic chlorophenols in a laboratory continuous-flow experiment. The results showed that the test compounds were concentrated in the device to an extent very closed to that in fish. The overall uptake rate constants (Ku) and average pollutant sampling rates (Rs) were similar for both SPMDs and goldfish. We conclude that the SPMDs can be used to estimate the levels of moderately hydrophobic chlorophenols in field water as an environmental monitor.

Animals↗

Planar gradient coil design by scaling the spatial frequencies of minimum-inductance current density.

Gradient coil inductance has been remarkably reduced by the minimum-inductance design technique, which minimizes the magnetic energy stored by the gradient coil. The planar gradient coil designed by this technique, however, often has poor magnetic field linearity. Scaling the spatial frequencies of the current density function derived by this method, the magnetic field linearity of the planar gradient coil can be greatly improved with a small sacrifice of gradient coil inductance. A figure of merit of the planar gradient coil has been found to be improved by scaling the spatial frequencies.

Equipment Design↗

[Preparation of leucine-methyl glutamate-glutamic acid copolymers].

The method for preparing leucine-methyl glutamate-glutamic acid copolymer was studied. In the first place benzyl glutamate and methyl glutamate were synthesized respectively. Then N-carboxy anhydrides (NCA) of leucine, benzyl glutamate or methyl glutamate were prepared in a closed container by phosgene-toluene solution method. After copolymerization the copolymers were debenzylated and demethylated by anhydrous hydrogen bromide. The free carboxyl group mole content in side chains of the copolymer was controlled by various standing periods following bubbling HBr. Analysis of infrared spectrogram and ultraviolet asorbance of copolymers indicated that this procedure resulted in the loss of almost all benzyl groups and some methyl groups.

Glutamic Acid↗