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Biomedical subjects

Y Zhu

Publications and source records attributed to Y Zhu.

At least 109 records · Page 6Linked to original sources
PubMed2001

Flash photolytic generation of ortho-quinone methide in aqueous solution and study of its chemistry in that medium.

Flash photolysis of o-hydroxybenzyl alcohol, o-hydroxybenzyl p-cyanophenyl ether, and (o-hydroxybenzyl)trimethylammonium iodide in aqueous perchloric acid and sodium hydroxide solutions, and in acetic acid and biphosphate ion buffers, produced o-quinone methide as a short-lived transient species that underwent hydration back to benzyl alcohol in hydrogen-ion catalyzed (k(H+) = 8.4 x 10(5) M(-1) s(-1)) and hydroxide-ion catalyzed (k(HO)- = 3.0 x 10(4) M(-1) s(-1)) reactions as well as an uncatalyzed (k(UC) = 2.6 x 10(2) s(-1)) process. The hydrogen-ion catalyzed reaction gave the solvent isotope effect k(H+)/k(D)+ = 0.42, whose inverse nature indicates that this process occurs by rapid and reversible equilibrium protonation of the carbonyl oxygen atom of the quinone methide, followed by rate-determining capture of the carbocation so produced by water. The magnitude of the rate constant of the uncatalyzed reaction, on the other hand, indicates that this process occurs by simple nucleophilic addition of water to the methylene group of the quinone methide. Decay of the quinone methide is also accelerated by acetic acid buffers through both acid- and base-catalyzed pathways, and quantitative analysis of the reaction products formed in these solutions shows that this acceleration is caused by nucleophilic reactions of acetate ion rather than by acetate ion assisted hydration. Bromide and thiocyanate ions also accelerate decay of the quinone methide through both hydrogen-ion catalyzed and uncatalyzed pathways, and the inverse nature of solvent isotope effects on the hydrogen-ion catalyzed reactions shows that these reactions also occur by rapid equilibrium protonation of the quinone methide carbonyl oxygen followed by rate-determining nucleophilic capture of the ensuing carbocation. Assignment of an encounter-controlled value to the rate constant for the rate-determining step of the thiocyanate reaction leads to pK(a) = -1.7 for the acidity constant of the carbonyl-protonated quinone methide.

Acetic Acid↗
PubMed2001

Cloning and characterization of PIMT, a protein with a methyltransferase domain, which interacts with and enhances nuclear receptor coactivator PRIP function.

The nuclear receptor coactivators participate in the transcriptional activation of specific genes by nuclear receptors. In this study, we report the isolation of a nuclear receptor coactivator-interacting protein from a human liver cDNA library by using the coactivator peroxisome proliferator-activated receptor-interacting protein (PRIP) (ASC2/AIB3/RAP250/NRC/TRBP) as bait in a yeast two-hybrid screen. Human PRIP-interacting protein cDNA has an ORF of 2,556 nucleotides, encodes a protein with 852 amino acids, and contains a 9-aa VVDAFCGVG methyltransferase motif I and an invariant GXXGXXI segment found in K-homology motifs of many RNA-binding proteins. The gene encoding this protein, designated PRIP-interacting protein with methyltransferase domain (PIMT), is localized on chromosome 8q11 and spans more than 40 kb. PIMT mRNA is ubiquitously expressed, with a high level of expression in heart, skeletal muscle, kidney, liver, and placenta. Using the immunofluorescence localization method, we found that PIMT and PRIP proteins appear colocalized in the nucleus. PIMT strongly interacts with PRIP under in vitro and in vivo conditions, and the PIMT-binding site on PRIP is in the region encompassing amino acids 773-927. PIMT binds S-adenosyl-l-methionine, the methyl donor for methyltransfer reaction, and it also binds RNA, suggesting that it is a putative RNA methyltransferase. PIMT enhances the transcriptional activity of peroxisome proliferator-activated receptor gamma and retinoid-X-receptor alpha, which is further stimulated by coexpression of PRIP, implying that PIMT is a component of nuclear receptor signal transduction apparatus acting through PRIP. Definitive identification of the specific substrate of PIMT and the role of this RNA-binding protein in transcriptional regulation remain to be determined.

Amino Acid Sequence↗
PubMed2001

High-performance affinity capture-removal of bacterial pyrogen from solutions.

Synthetic peptide S3delta has high affinity for bacterial endotoxin or lipopolysaccharide (LPS). Under tested conditions of pH 5-9 and 0-0.4 M NaCl, the affinity constant, KD ranged from 2.10(-6) to 2 x 10(-9) M(-1). A novel affinity matrix based on peptide S3delta was developed for removal of LPS from solutions such as: water; buffers with a wide range of ionic strength and pH; medium for cell culture; and protein solutions under optimized conditions. At a starting LPS of approximately 100 EU/ml, a post-purification level below 0.005 EU/ml was achieved.

Amino Acid Sequence↗
PubMed2001

Viscosity of interfacial water.

The effective shear viscosity and frequency-dependent dynamic oscillatory shear spectra of water containing monovalent or divalent ions (ionic strength 25 mM), confined between mica crystals at 1-2 water molecules thickness, oscillated with twist angle with the period expected for the pseudohexagonal surface lattice. The effective viscosity varied by orders of magnitude as the twist angle was changed. Confinement appeared to imprint lateral spatial correlation on the ultrathin liquid, the more so the better the confining lattices were aligned, but the oft-proposed "ice structure" was not observed dynamically.

Viscosity↗
PubMed2001

Rate-dependent slip of Newtonian liquid at smooth surfaces.

Newtonian fluids were placed between molecularly smooth surfaces whose spacing was vibrated at spacings where the fluid responded as a continuum. Hydrodynamic forces agreed with predictions from the no-slip boundary condition only provided that flow rate (peak velocity normalized by spacing) was low, but implied partial slip when it exceeded a critical level, different in different systems, correlated with contact angle (surface wettability). With increasing flow rate and partially wetted surfaces, hydrodynamic forces became up to 2-4 orders of magnitude less than expected by assuming the no-slip boundary condition that is commonly stated in textbooks.

Journal Article↗
PubMed2001

Bonding mechanism and atomic geometry of an ordered hydroxyl overlayer on Pt(111).

Exposing water to a (2 x 2)-O precovered Pt(111) surface at 100 K and subsequently annealing at 155 K led to the formation of a well-ordered (square root 3 x square root 3)R30 degrees overlayer. The structure of this overlayer is determined by DFT and full dynamical LEED calculations. There are two O containing groups per (square root 3 x square root 3)R30 degrees unit cell and both occupy near on-top positions with a Pt-O bond length of (2.11 +/- 0.04) A. DFT calculations determined the hydrogen positions of the OH species and clearly indicate hydrogen bonds between the neighboring adsorbed OH groups whose interaction is mainly of electrostatic nature. A theoretical comparison with H(2)O shows the hybridization of OH on Pt(111) to be sp(3).

Journal Article↗
PubMed2001

An essential role of the JAK-STAT pathway in ischemic preconditioning.

The goal of this study was to determine the role of the Janus tyrosine kinase (JAK)-signal transducers and activators of transcription (STAT) pathway in the late phase of ischemic preconditioning (PC). A total of 230 mice were used. At 5 min after ischemic PC (induced with six cycles of 4-min coronary occlusion/4-min reperfusion), immunoprecipitation with anti-phosphotyrosine (anti-pTyr) antibodies followed by immunoblotting with anti-JAK antibodies revealed increased tyrosine phosphorylation of JAK1 (+257 +/- 53%) and JAK2 (+238 +/- 35%), indicating rapid activation of these two kinases. Similar results were obtained by immunoblotting with anti-pTyr-JAK1 and anti-pTyr-JAK2 antibodies. Western analysis with anti-pTyr-STAT antibodies demonstrated a marked increase in nuclear pTyr-STAT1 (+301 +/- 61%) and pTyr-STAT3 (+253 +/- 60%) 30 min after ischemic PC, which was associated with redistribution of STAT1 and STAT3 from the cytosolic to the nuclear fraction and with an increase in STAT1 and STAT3 gamma-IFN activation site DNA-binding activity (+606 +/- 64%), indicating activation of STAT1 and STAT3. No nuclear translocation or tyrosine phosphorylation of STAT2, STAT4, STAT5A, STAT5B, or STAT6 was observed. Pretreatment with the JAK inhibitor AG-490 20 min before the six occlusion/reperfusion cycles blocked the enhanced tyrosine phosphorylation of JAK1 and JAK2 and the increased tyrosine phosphorylation, nuclear translocation, and enhanced DNA-binding activity of STAT1 and STAT3. The same dose of AG-490 abrogated the protection against myocardial infarction and the concomitant up-regulation of inducible NO synthase (iNOS) protein and activity observed 24 h after ischemic PC. Taken together, these results demonstrate that ischemic PC induces isoform-selective activation of JAK1, JAK2, STAT1, and STAT3, and that ablation of this response impedes the up-regulation of iNOS and the concurrent acquisition of ischemic tolerance. This study demonstrates that the JAK-STAT pathway plays an essential role in the development of late PC. The results reveal a signaling mechanism that underlies the transcriptional up-regulation of the cardiac iNOS gene and the adaptation of the heart to ischemic stress.

Animals↗
PubMed2001

Removal of ECG interference from the EEG recordings in small animals using independent component analysis.

In experiments involving small animals, the electroencephalogram (EEG) recorded during severe injury and accompanying resuscitation exhibit the strong presence of electrocardiogram (ECG). For improved quantitative EEG (qEEG) analysis, it is therefore imperative to remove ECG interference from EEG. In this paper, we validate the use of independent component analysis (ICA) to effectively suppress the interference of ECG from EEG recordings during normal activity, asphyxia and recovery following asphyxia. Two channels of EEG from five rats were recorded continuously for 2 h. Simultaneous recording of one channel ECG was also made. Epochs of 4 s and 1 min were selected from baseline, asphyxia and recovery (every 10 min) and their independent components and power spectra were calculated. The improvement in normalized power spectrum of EEG obtained for all animals was 7.71+/-3.63 db at the 3rd minute of recovery and dropped to 1.15+/-0.60 db at 63rd minute. The application of ICA has been particularly useful when the power of EEG is low, such as that observed during early brain hypoxic-asphyxic injury. The method is also useful in situations where accurate indications of EEG signal power and frequency content are needed.

Algorithms↗
PubMed2001

Plasminogen activator inhibitor type 1 enhances neointima formation after oxidative vascular injury in atherosclerosis-prone mice.

BACKGROUND: Plasminogen activator inhibitor type 1 (PAI-1) inhibits neointima formation after vascular injury. Hyperlipidemia modulates the expression of multiple genes, however, and the effects of PAI-1 on the arterial response to injury under hyperlipidemic conditions are unknown. The purpose of this study was to examine the impact of PAI-1 on intimal hyperplasia and other vascular changes that develop after arterial injury in apolipoprotein E-deficient (apoE(-/-)) mice. METHODS AND RESULTS: Ferric chloride injury of the midportion of the common carotid arteries of apoE(-/-) mice (n=22) induced formation of a neointima that contained smooth muscle cells, foam cells, neutral lipid, tissue factor, and von Willebrand factor. Interactions between vascular injury and apolipoprotein E deficiency were strongly synergistic; either stimulus alone was insufficient to induce significant neointima formation. Mean intima/media ratios were significantly greater (P<0.03) in apoE(-/-), PAI-1(+/+) mice (5.6+/-1.8, n=12) than in apoE(-/-), PAI-1(-/-) mice (1.2+/-0.55, n=12), as were the percentages of bromodeoxyuridine-positive cells in the intima and media (P<0.03). Transiently occlusive (<48 hours) and nonocclusive mural thrombi persisted longer in apoE(-/-), PAI-1(+/+) mice than in apoE(-/-), PAI-1(-/-) mice. CONCLUSIONS: In atherosclerosis-prone mice, PAI-1 promotes neointima formation after oxidative vascular injury. The apparent hyperlipidemia-dependent effect of PAI-1 may be mediated by its capacity to inhibit the clearance of platelet-fibrin thrombi that can deliver growth factors to the blood vessel wall or be incorporated into developing vascular lesions. Alternatively, hyperlipidemia may alter the pattern of gene expression in the blood vessel wall to enhance potential effects of PAI-1 on antiproliferative processes, such as transforming growth factor-beta activation and apoptosis.

Animals↗
PubMed2001

[Mutation analysis of tumor suppressor gene PTEN in bone and soft tissue tumors].

OBJECTIVE: To study tumor suppressor gene PTEN's role in the tumorigenesis of bone and soft tissue tumors. METHODS: Exon 4 approximately 9 of PTEN were amplified by polymerase chain reaction (PCR) for homozygous deletions in 110 tumor samples of patients with bone and soft tissue tumors, 3 tumor cell lines. PTEN mutations were detected by a combination of single strand conformation polymorphism (SSCP) analysis and DNA sequencing. Normal tissue beside tumors was used as control. RESULTS: There were always products of amplified exon 4 approximately 9 of PTEN, so no homozygous deletions existed. No homozygous deletions presented as variant band was not found by SSCP analysis. Two kinds of band type appeared in exon 8. A g/t polymorphism was found 32 bp from the splice donor site of intron 8 in six tumors. CONCLUSIONS: At the level of DNA, PTEN mutations do not play major roles in the tumorigenesis of bone and soft tissue tumors. The polymorphism of PTEN gene exists in Chinese people.

Asian People↗
PubMed2001

Sucrase-isomaltase gene transcription requires the hepatocyte nuclear factor-1 (HNF-1) regulatory element and is regulated by the ratio of HNF-1 alpha to HNF-1 beta.

The mouse sucrase-isomaltase (SI) gene is an enterocyte-specific gene expressed in a complex developmental pattern. We previously reported that a short, evolutionarily conserved gene promoter regulates developmental expression of SI in mouse small intestine. Herein, we investigated the role of a hepatocyte nuclear factor-1 (HNF-1) cis-acting element to regulate SI gene expression in vivo. Transgenic SI gene constructs with a mutated HNF-1 element (SIF3) revealed a strong reduction in promoter activity in comparison with a wild-type construct in mice and during Caco-2 cell differentiation. Nuclear proteins isolated from enterocytes showed increased binding of the HNF-1 alpha complex with a concomitant decrease in the HNF-1 beta-containing complex to the SIF3 element both during the suckling-weaning developmental transition and Caco-2 cell differentiation. These changes coincided with a strong induction of SI gene transcription. In transfection experiments, HNF-1 alpha activated the SI promoter via the SIF3 element, and co-expression of HNF-1 beta impaired this transcriptional activation. These findings demonstrate the essential role of the HNF-1 regulatory element to support SI gene transcription in vivo and suggest that the ratio of HNF-1 alpha to HNF-1 beta plays a role in the transcriptional activity of this gene during intestinal development.

Animals↗
PubMed2001

Binding of GGA2 to the lysosomal enzyme sorting motif of the mannose 6-phosphate receptor.

The GGAs are a multidomain protein family implicated in protein trafficking between the Golgi and endosomes. Here, the VHS domain of GGA2 was shown to bind to the acidic cluster-dileucine motif in the cytoplasmic tail of the cation-independent mannose 6-phosphate receptor (CI-MPR). Receptors with mutations in this motif were defective in lysosomal enzyme sorting. The hinge domain of GGA2 bound clathrin, suggesting that GGA2 could be a link between cargo molecules and clathrin-coated vesicle assembly. Thus, GGA2 binding to the CI-MPR is important for lysosomal enzyme targeting.

Adaptor Proteins, Vesicular Transport↗
PubMed2001

Suppression of a sialyltransferase by antisense DNA reduces invasiveness of human colon cancer cells in vitro.

Transfer of terminal alpha 2,6-linked sialic acids to N-glycans is catalyzed by beta-galactoside alpha 2,6-sialyltransferase (ST6Gal I). Expression of ST6Gal I and its products is reportedly increased in colon cancers. To investigate directly the functional effects of ST6Gal I expression, human colon cancer (HT29) cells were transfected with specific antisense DNA. ST6Gal I mRNA and protein were virtually undetectable in six strains of transfected HT29 cells. ST6Gal activity was reduced to 14% of control (P<0.005) in transfected cells. Expression of terminal alpha 2,6- and alpha 2,3-linked sialic acids, and unmasked N-acetyllactosamine oligosaccharides, respectively, was assessed using flow cytometry and fluoresceinated Sambucus nigra, Maackia amurensis and Erythrina cristagalli lectins. Results indicated a major reduction in expression of alpha 2,6-linked sialic acids and counterbalancing increase in unmasked N-acetyllactosamines in antisense DNA-transfected cells, without altered expression of alpha 2,3-linked sialic acids or ganglioside profiles. The ability of transfected cells to form colonies in soft agar and to invade extracellular matrix material (Matrigel), respectively, in vitro was reduced by approx. 98% (P<0.0001) and more than 3-fold (P<0.005) compared to parental HT29 cells. These results indicate that N-glycans bearing terminal alpha 2,6-linked sialic acids may enhance the invasive potential of colon cancer cells.

Cell Aggregation↗
PubMed2001

A facile and effective synthesis of alpha-(1-->6)-linked mannose di-, tri-, tetra-, hexa-, octa-, and dodecasaccharides, and beta-(1-->6)-linked glucose di-, tri-, tetra-, hexa-, and octasaccharides using sugar trichloroacetimidates as the donors and unprotected or partially protected glycosides as the acceptors.

Reaction of 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl trichloroimidate with allyl alpha-D-mannopyranoside in the presence of TMSOTf selectively gave allyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-alpha-D-mannopyranoside through an orthoester intermediate. Benzoylation of 3, followed by deallylation, and then trichloroimidation afforded the disaccharide donor 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroimidate, while benzoylation of 3 followed by selective removal of acetyl groups yielded the disaccharide acceptor allyl alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranoside. Coupling of 5 with 6 gave the tetrasaccharide allyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranoside, which were converted into the tetrasaccharide donor 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroimdate and the tetrasaccharide acceptor allyl alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranoside, respectively, by the same strategies as used for conversion of 3 into 5 and 6. Condensation of 5 with 13 gave the hexasaccharide 14, while condensation of 12 with 13 gave the octasaccharide 17. Dodecasaccharide 21 was obtained by the coupling of 12 with the octasaccharide acceptor 20. Similar strategies were used for the syntheses of beta-(1-->6)-linked glucose di-, tri-, tetra-, hexa-, and octamers. Deprotection of the oligosaccharides in ammonia-saturated methanol yielded the free alpha-(1-->6)-linked mannosyl and beta-(1-->6)-linked glucosyl oligomers.

Acetamides↗
PubMed2001

Convergent synthesis of peptide conjugates using dehydroalanines for chemoselective ligations.

[reaction: see text]. Protein and peptide conjugates such as glycopeptides, prenylated peptides, and lipopeptides play essential roles in biology. A rapid and convergent entry into a variety of these compounds is described. The methodology involves the introduction of a dehydroalanine into peptides and subsequent chemoselective conjugate addition of an appropriate thiolate nucleophile, including farnesylthiolate or thioglycosides.

Alanine↗
PubMed2001

Ablation of NF1 function in neurons induces abnormal development of cerebral cortex and reactive gliosis in the brain.

Neurofibromatosis type 1 (NF1) is a prevalent genetic disorder that affects growth properties of neural-crest-derived cell populations. In addition, approximately one-half of NF1 patients exhibit learning disabilities. To characterize NF1 function both in vitro and in vivo, we circumvent the embryonic lethality of NF1 null mouse embryos by generating a conditional mutation in the NF1 gene using Cre/loxP technology. Introduction of a Synapsin I promoter driven Cre transgenic mouse strain into the conditional NF1 background has ablated NF1 function in most differentiated neuronal populations. These mice have abnormal development of the cerebral cortex, which suggests that NF1 has an indispensable role in this aspect of CNS development. Furthermore, although they are tumor free, these mice display extensive astrogliosis in the absence of conspicuous neurodegeneration or microgliosis. These results indicate that NF1-deficient neurons are capable of inducing reactive astrogliosis via a non-cell autonomous mechanism.

Alleles↗
PubMed2001

The electrochemically induced conformational transition of disulfides in bovine serum albumin studied by thin layer circular dichroism spectroelectrochemistry.

The conformational transition of disulfides in bovine serum albumin (BSA) induced by electrochemical redox reaction of disulfides were monitored by in-situ circular dichroism (CD) spectroelectrochemistry, with a long optical path thin layer cell and analyzed by a singular value decomposition least square (SVDLS) method. Electrochemical reduction of disulfides drives the left-handed conformation of disulfides changed into the right-handed. At open circuit, eight of the 17 disulfides were of left-handed conformation. Four of the 17 disulfides took part in the electrochemical reduction with an EC mechanism. Only one-fourth of the reduced disulfides returned to left-handed conformation in the re-oxidation process. Some parameters of the electrochemical reduction process, i.e. the number of electrons transferred and electron transfer coefficient, n = 8, alpha n = 0.15, apparent formal potential, E1(0') = -0.65(+/-0.01) V, standard heterogeneous electron transfer rate constant, k1(0) = (2.84 +/- 0.14) x 10(-5) cm s(-1) and chemical reaction equilibrium constant, Kc = (5.13 +/- 0.12) x 10(-2), were also obtained by double logarithmic analysis based on the near-UV absorption spectra with applied potentials.

Algorithms↗