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Biomedical subjects

Z Yu

Publications and source records attributed to Z Yu.

At least 37 records · Page 2Linked to original sources

[Acetylcholine and A23187 stimulate epinephrine secretion of isolated rat adrenal medulla cells in vitro].

Acetylcholine and A23187, two acetylcholinergic agonists, were used to induce the stimulus-secretion coupling of the isolated rat adrenal medulla cells. Morphometry of numerical density and electron probe X-ray quantitative microanalysis were adopted to measure the alterations in the number and calcium content of chromaffin granules in adrenal medulla cells during the agonist treatment. Secretion of epinephrine was detected by the high performance liquid chromatography (HPLC). It was found that the Ca content of the isolated rat adrenal medulla chromaffin granules dropped significantly after a 10-min incubation with the agonists, whereas the number of chromaffin granule decreased slowly during the agonist treatment and the epinephrine concentration raised significantly after a 20-min incubation with the agonists. The decrease of granule Ca content occurred earlier than the increase of epinephrine concentration, suggesting that the calcium released from chromaffin granule may in part account for the induced cell secretion.

Acetylcholine↗

Soluble epoxide hydrolase regulates hydrolysis of vasoactive epoxyeicosatrienoic acids.

The cytochrome P450-derived epoxyeicosatrienoic acids (EETs) have potent effects on renal vascular reactivity and tubular sodium and water transport; however, the role of these eicosanoids in the pathogenesis of hypertension is controversial. The current study examined the hydrolysis of the EETs to the corresponding dihydroxyeicosatrienoic acids (DHETs) as a mechanism for regulation of EET activity and blood pressure. EET hydrolysis was increased 5- to 54-fold in renal cortical S9 fractions from the spontaneously hypertensive rat (SHR) relative to the normotensive Wistar-Kyoto (WKY) rat. This increase was most significant for the 14,15-EET regioisomer, and there was a clear preference for hydrolysis of 14, 15-EET over the 8,9- and 11,12-EETs. Increased EET hydrolysis was consistent with increased expression of soluble epoxide hydrolase (sEH) in the SHR renal microsomes and cytosol relative to the WKY samples. The urinary excretion of 14,15-DHET was 2.6-fold higher in the SHR than in the WKY rat, confirming increased EET hydrolysis in the SHR in vivo. Blood pressure was decreased 22+/-4 mm Hg (P:<0.01) 6 hours after treatment of SHRs with the selective sEH inhibitor N:, N:'-dicyclohexylurea; this treatment had no effect on blood pressure in the WKY rat. These studies identify sEH as a novel therapeutic target for control of blood pressure. The identification of a potent and selective inhibitor of EET hydrolysis will be invaluable in separating the vascular effects of the EET and DHET eicosanoids.

8,11,14-Eicosatrienoic Acid↗

Binding interactions of vancomycin tracers with a bacterial cell wall peptidoglycan analogue.

Binding interactions between several vancomycin tracers and (N,N'-diacetyl)KDADA in solution were evaluated in a competition format using a surface plasmon resonance instrument. Tracers derivatized from the carboxy terminus or the N-vancosaminyl sugar moiety of vancomycin bind the peptide with an affinity similar to that of underivatized vancomycin. In contrast, N-methylleucyl derivatized vancomycin tracers bind the peptide with a reduced affinity relative to vancomycin.

Anti-Bacterial Agents↗

Transposon diversity in Arabidopsis thaliana.

Recent availability of extensive genome sequence information offers new opportunities to analyze genome organization, including transposon diversity and accumulation, at a level of resolution that was previously unattainable. In this report, we used sequence similarity search and analysis protocols to perform a fine-scale analysis of a large sample ( approximately 17.2 Mb) of the Arabidopsis thaliana (Columbia) genome for transposons. Consistent with previous studies, we report that the A. thaliana genome harbors diverse representatives of most known superfamilies of transposons. However, our survey reveals a higher density of transposons of which over one-fourth could be classified into a single novel transposon family designated as Basho, which appears unrelated to any previously known superfamily. We have also identified putative transposase-coding ORFs for miniature inverted-repeat transposable elements (MITEs), providing clues into the mechanism of mobility and origins of the most abundant transposons associated with plant genes. In addition, we provide evidence that most mined transposons have a clear distribution preference for A + T-rich sequences and show that structural variation for many mined transposons is partly due to interelement recombination. Taken together, these findings further underscore the complexity of transposons within the compact genome of A. thaliana.

Amino Acid Sequence↗

Human G protein gamma(11) and gamma(14) subtypes define a new functional subclass.

The mammalian gamma subunit family consists of a minimum of 12 members. Analysis of the amino acid sequence conservation suggests that the gamma subunit family can be divided into three distinct subclasses. The division of the gamma subunit family into these classes is based not only on amino acid homology, but also to some extent on functional similarities. In the present study, two new members of the gamma subunit family, the gamma(11) and gamma(14) subunits, are identified and characterized in terms of their expression and function. The gamma(11) and gamma(14) subunits are most closely related to the gamma(1) subunit and share similar biochemical properties, suggesting their inclusion in class I. However, despite their close phylogenetic relationship and similar biochemical properties, the gamma(1), gamma(11), and gamma(14) subunits exhibit very distinct expression patterns, suggesting that class I should be further subdivided and that the signaling functions of each subgroup are distinct. In this regard, the gamma(11) and gamma(14) subunits represent a new subgroup of farnesylated gamma subunits that are expressed outside the retina and have functions other than phototransduction.

Amino Acid Sequence↗

Direct analysis of artemisinin in plasma and saliva using coupled-column high-performance liquid chromatography with a restricted-access material pre-column.

A previously established HPLC system with post-column derivatization for the analysis of artemisinin was coupled to an ADS (alkyl-diol silica) pre-column, allowing direct and repetitive injection of protein-rich fluids such as plasma. The limit of quantitation for 100 microl of plasma was 10 ng/ml (CV=10.5%) while concentrations down to 2 ng/ml could be quantified for 1.00 ml saliva samples (CV=11.1%). The system was linear in the tested range of 10-2000 ng/ml for plasma and 2-240 ng/ml for saliva samples, respectively. This paper introduces coupled column HPLC as a simplified method for the routine analysis of artemisinin in biological fluids.

Antimalarials↗

The dominant negative LQT2 mutation A561V reduces wild-type HERG expression.

HERG(1) K(+) channel mutations are responsible for one form of dominantly inherited long QT syndrome (LQT). Some LQT mutations exert a dominant negative effect on wild-type current expression. To investigate mechanisms of dominant-negative behavior, we co-expressed wild-type HERG with the A561V mutant in mammalian cells. Transfection with various cDNA ratios produced HERG K(+) current densities that approached a predicted binomial distribution where mutant and wild-type subunits co-assemble in a tetramer with nearly complete dominance. Using C terminus myc-tagged wild-type HERG we specifically followed the mutant's effect on full-length wild-type HERG protein expression. Co-expression with A561V reduced the abundance of full-length wild-type HERG protein comparable to the current reduction. Reduction of wild-type protein was due to decreased synthesis and increased turnover. Conditions facilitating protein folding (growth at 30 degrees C, or in 10% glycerol) resulted in partial rescue from the dominant effect, as did the 26 S proteosome inhibitor ALLN. Thus, for A561V, dominant negative effects result from assembly of wild-type subunits with mutant very early in production leading to rapid recognition of mutant channels and targeting for proteolysis. These results establish protein misfolding, cellular proofreading, and bystander involvement as contributing mechanisms for dominant effects in LQT2.

Animals↗

P(MeNCH2CH2)3N: an efficient catalyst for the desilylation of tert-butyldimethylsilyl ethers

tert-Butyldimethylsilyl (TBDMS) ethers of primary, secondary, and tertiary alcohols and phenolic TBDMS ethers are desilylated to their corresponding alcohols and phenols, respectively, in DMSO, at 80 degrees C, in 68-94% yield in the presence of 0.2-0.4 equiv of P(MeNCH2CH2)3N. Using P(i-PrNCH2-CH2)3N as the catalyst, 85-97% yields of desilylated alcohols were obtained from TBDMS ethers of 1-octanol, 2-phenoxyethanol, and racemic alpha-phenyl ethanol. These are the first examples of desilylations of silyl ethers catalyzed by nonionic bases. Both catalysts were much less effective for the desilylation of tert-butyldiphenylsilyl (TBDPS) ethers (22-45% yield) under the same conditions as used for TBDMS ethers. Possible pathways involving nucleophilic attack of the anion of the solvent molecule (generated by the catalyst) at the Si-O bond of silyl ether or a prior activation of the silyl ether by the catalyst via a P-Si interaction followed by nucleophilic attack of the solvent anion are proposed on the basis of 1H and 31P NMR experimental data.

Journal Article↗

HER-2/neu blocks tumor necrosis factor-induced apoptosis via the Akt/NF-kappaB pathway.

Overexpression of HER-2/neu correlates with poor survival of breast and ovarian cancer patients and induces resistance to tumor necrosis factor (TNF), which causes cancer cells to escape from host immune defenses. The mechanism of HER-2/neu-induced TNF resistance is unknown. Here we report that HER-2/neu activates Akt and NF-kappaB without extracellular stimulation. Blocking of the Akt pathway by a dominant-negative Akt sensitizes the HER-2/neu-overexpressing cells to TNF-induced apoptosis and inhibits IkappaB kinases, IkappaB phosphorylation, and NF-kappaB activation. Our results suggested that HER-2/neu constitutively activates the Akt/NF-kappaB anti-apoptotic cascade to confer resistance to TNF on cancer cells and reduce host defenses against neoplasia.

3T3 Cells↗

Abnormal expression of hepatoma specific gamma-glutamyl transferase and alteration of gamma-glutamyl transferase gene methylation status in patients with hepatocellular carcinoma.

BACKGROUND: Hepatoma specific gamma-glutamyl transferase (HS-GGT) bands were expressed in the development of hepatocellular carcinoma (HCC) and were associated with a high incidence of HCC diagnosis. The objectives of this study were to determine the levels of HS-GGT quantitatively in the sera of patients with different liver diseases. The methylational status of GGT gene CCGG sites was analyzed in hepatoma tissues. METHODS: The HS-GGT concentrations were quantitatively analyzed in the sera of 156 HCC patients and others with liver diseases or extrahepatic tumors. In 20 hepatoma tissues, the GGT enzyme proteins were purified, the activities of GGTs of different molecular form were examined, total RNAs were extracted and amplified by using a nested polymerase chain reaction (PCR) assay, and the methylational status of CCGG site (M3) in the 5'-noncoding region of GGT genes was investigated with the restriction enzyme Hpa II. RESULTS: Total GGT activities in patients with liver diseases and extrahepatic tumors were abnormally increased. The levels of serum HS-GGT were significantly elevated (P < 0.001) in the HCC group; the incidence of HS-GGT over 5.5 IU/L was 86% in HCC patients and less than 3% in patients with other diseases. From liver cancer to distal noncancerous tissues, an increasing tendency (P < 0.05) of total RNA concentrations was found; the frequencies of amplified fragment and hypomethylated M3 site of GGT genes were 100% and 75% in HCC, 85% and 55% in paracancerous tissues, and 75% and 50% in noncancerous tissues, respectively. An inverse correlation was found between methylational degrees of GGT genes and expression levels of GGT. CONCLUSIONS: The abnormal alteration of serum HS-GGT level is a sensitive tumor marker for HCC diagnosis or differentiation, and the overexpression of GGT in HCC may be related to the hypomethylational status of CCGG sites of GGT genes.

Adult↗

Mass spectrometric study of six cyclic esters

A series of cyclic esters, which are optically active as a consequence of their helical structures, were synthesized to investigate the relationships between their structures and their optical activities. This paper reports the electron impact fragmentation mechanisms of these six cyclic esters. Accurate mass measurements and mass analyzed ion kinetic energy spectrometry confirmed fragmentation patterns. The stability of the fragment ions has a great influence on the fragmentation pathways, but no correlation with the optical activity was found.

Journal Article↗

Two novel myogenic factors identified and isolated by sequential isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

A number of environmental factors were used experimentally to enhance myogenesis during muscle regeneration; however, many hormones and growth factors have been shown to have the ability to increase the rate of satellite cell division, but they only work on satellite cells that are already active in many animal experiments. Recently, the crushed muscle extract (CME) of rats was found to be able to trigger dormant adult rat satellite cells to re-enter the cell mytogenic cycle; however, the identity of the active factors present in rat CME remains unknown. In the present study, the CME was fractionated by the strategy of sequential isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) coupled with functional analysis by myoblast culture. Two satellite cell-specific myogenic factors were identified and purified from CME by this strategy. One of the factors has a molecular mass of around 7 kDa and another about 39 kDa. The factor of 39 kDa could be retained in heparin-Sepharose column and eluted with phosphate-buffered saline (PBS) containing 1 M NaCl, but the 7 kDa factor did not bind to the heparin column. These two purified myogenic factors could synergistically trigger the proliferation and differentiation of dormant satellite cells, whose progenies subsequently fuse in vitro, or fuse to pre-existing partially damaged muscle fibers to form full repair of the damaged muscle fibers or to form new myotubes to replace the completely damaged muscle fibers during the cascade of muscle healing and regeneration in vivo. The identities of these two myogenic factors are under study.

Animals↗

Electron impact fragmentation mechanisms of some cyclic esters with helical structures

The electron impact mass spectra of several cyclic esters with helical structures have been studied. Their fragmentation pathways were proposed and confirmed by mass-analyzed ion kinetic energy (MIKE) and high-resolution data. In general, the dominant fragmentation pathways in the spectra of these compounds originate from a alpha-cleavage with loss of a hydrogen or methyl group. The difference between hydrogen and methyl group loss greatly affects the subsequent fragmentations. Although, due to their helicity, these cyclic esters are optically active no stereo-related fragmentation pathway was observed. Copyright 2000 John Wiley & Sons, Ltd.

Journal Article↗

An association study of polymorphisms in the alpha-antichymotrypsin gene for Alzheimer disease in Han-Chinese.

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and DNA sequencing were employed to screen the coding region of the alpha-antichymotrypsin (AACT) gene in Han-Chinese population for polymorphism possibly associated with Alzheimer's disease (AD). Consequently, seven polymorphic sites including 25A>G, 39G>A, 370C>T, 662T>G, 892C>T, 923T>C and 1332A>G were detected. Of them, the 25A>G was reported previously and the others are all novel. We subsequently focused on the 25A>G and the 39G>A polymorphism that were of interest to us and conducted an association study of them by another scanning of 246 controls that matched the AD patients. Statistic test showed that both genotype (p=0.0378, Fisher's exact, two tailed) and allele frequency (p=0.0382, Fisher's exact, two tailed) of 39G>A are different between AD patients and the controls. As for 25A>G, lain only the heterozygous genotype A/G associates with AD (p=0.0220, chi(2)), but not the A allele frequency (p=0.1141, chi(2)).

Aged↗

Identification of genes responsive to BPDE treatment in HeLa cells using cDNA expression assays.

Genotoxic stresses induce cellular responses that can be observed at the level of gene expression. We have studied changes in gene expression following BPDE exposure in HeLa cells by using a cDNA expression array of 597 human genes. After a 53-hr exposure to 0.4 microM BPDE, nine genes were upregulated. The protein products of these genes are: fos-related antigen 2, apoptotic cysteine protease MCH4, DB1 (zinc finger protein 91), transcription factor ETR103, integrin alpha, interleukin-4, interleukin-6, 23-kDa highly basic protein, and ribosomal protein S9. We observed the downregulation of gene expression of three genes: heat-shock protein 27, DNA-binding protein TAX, and NADH-ubiquinone oxidoreductase B18 subunit. These results suggest unknown functions or regulatory circuits for several of the responsive genes and demonstrate the complexity of cellular responses to genotoxic insults.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

Application of surface plasmon resonance toward studies of low-molecular-weight antigen-antibody binding interactions.

Methods for studying low-molecular-weight antigen-antibody binding interactions using surface plasmon resonance detection are presented. The experimental parameters most relevant to studies of low-molecular-weight antigen-antibody binding interactions are discussed. Direct kinetic analysis of the binding interactions is most informative, providing both apparent association and dissociation rate constants from which equilibrium constants can be calculated. Equilibrium analysis, including steady-state and solution affinity studies, offers an alternative approach to direct kinetic analysis when knowledge of the individual kinetic rate constants is not required or difficult to determine. The various methods are illustrated by studies of an anti-T(4) Fab fragment binding interaction with several thyroxine analogs. The methods utilized were dependent on the affinity of the interaction. The high-affinity anti-T(4) Fab fragment/l-T(4) binding interaction was evaluated using direct kinetic analysis. An intermediate affinity anti-T(4) Fab fragment/l-T(3) binding interaction was evaluated using a combination of direct kinetic analysis, steady-state analysis, and solution affinity analysis. The relatively weak anti-T(4) Fab fragment/l-T(2) binding interaction was evaluated using steady-state and solution affinity analysis protocols. Several thyroxine tracers that could not be immobilized to a biosensor surface were also evaluated via the solution affinity format. In cases where a given binding interaction was examined using multiple methods the results were comparable.

Animals↗

Change of coagulation factor VIII and antithrombin III activity in bank-stored blood.

Coagulation factor VIII and antithrombin III activity were detected in 15 health donors. It was found that antithrombin III activity decreased obviously 12 h after blood drawing. It lost 56% of the activity at the 3rd day, and 70% of the activity at the 7th day. FVIII:c showed no obvious change after 24 h, until the 3rd day. It lost 40%-60% of the activity after 36 h and was reduced to the 30% of the original activity at the 5th day. Our results suggested that at the 3rd day coagulation factor VIII of bank-stored blood can be used to replenish antithrombin III, while bank-stored blood in one day can be used to replenish F VIII.

Adolescent↗