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Biomedical subjects

Z Wu

Publications and source records attributed to Z Wu.

At least 541 records · Page 30Linked to original sources
PubMed1997

[Mitomycin C concentrations in rabbit ocular tissues after topical administration during glaucoma filtration surgery].

Using high-performance liquid chromatography, we measured Mitomycin C (MMC) concentrations in conjunctiva, sclera and aqueous of 22 rabbit eyes after single topical administration of 0.2 mg.ml-1 MMC during glaucoma filtration surgery. The peaks of MMC concentrations in conjunctiva, sclera, aqueous were 2.01 micrograms.g-1, 2.95 micrograms.g-1 and 0.16 microgram.ml-1 with half life of 0.63, 0.35 and 0.84 hours respectively. Irrigating the ocular surface with 20 ml of normal saline after MMC application reduced the peak drug concentration to 1/8 in conjunctiva, to 1/5 in sclera and to 1/3 in aqueous. The results showed that the MMC concentrations in conjunctiva and sclera were well above the ID50 of rabbit subconjunctival fibroblast (0.1 microgram.ml-1), and the concentration in aqueous was well below the level known to cause endothelium toxicity (approximately 0.2 mg.ml-1) and retinal toxicity (> 1.3 micrograms.ml-1). Therefore, 0.2 mg.ml-1 MMC can inhibit subconjunctival fibroblast effectively and has no side effect on visual function.

Administration, Topical↗
PubMed1996

Effect of migration and related environmental changes on serum lipid levels in southwestern Chinese men.

To determine migration-related differences in serum lipid levels and to estimate the contribution of environmental factors to these differences, the authors conducted a cross-sectional study among 1,222 residents of southwestern China in 1988 and 1989: 521 Yi farmers living in a remote rural area, 340 Yi migrants to an urban setting, and 361 Han people living in the same urban setting. Serum total and high density lipoprotein cholesterol and serum triglyceride were measured after a 14-hour fast. Information on age, cigarette smoking, alcohol consumption, and physical activity was obtained by questionnaire. Diet was assessed by use of three 24-hour diet recalls. The Yi farmers had the lowest age-adjusted serum lipid levels among the three groups, white the Yi migrants and their Han counterparts had similar lipid levels. After adjustment for age, body mass index, cigarette smoking, alcohol drinking, and dietary nutrient intake, the difference in serum lipid profile between the Yi migrants and the Yi farmers was substantially reduced. These results indicate that rural-urban migration is associated with higher serum lipid levels. This migration-related difference in serum lipids can be partially explained by changes in lifestyle and dietary nutrient intake. The authors conclude that environmental factors are important determinants of population serum lipid levels in southwestern Chinese.

Adolescent↗
PubMed1996

Biochemical and biophysical properties of the core-binding factor alpha2 (AML1) DNA-binding domain.

The Runt domain is the DNA-binding domain defining a small family of transcription factors that are involved in important developmental processes. Developmental pathways controlled by Runt domain proteins include sex determination, neurogenesis, segmentation, and eye development in Drosophila and hematopoiesis in mammals. In addition to binding DNA, the Runt domain also mediates heterodimerization with another subunit called the core-binding factor beta (CBFbeta) subunit. In this study we overexpress the Runt domain from the mouse CBFalpha2 (AML1) protein in Escherichia coli, and purify it from the insoluble fraction. We determine the equilibrium constants for Runt domain binding to two different DNA sequences by surface plasmon resonance technology. Circular dichroism spectroscopy demonstrates that the Runt domain is a folded beta-domain with essentially no alpha-helical content. The single tryptophan residue in the CBFalpha2 Runt domain at amino acid 79 is shown by tryptophan fluorescence spectroscopy to reside in a polar environment. Finally, we demonstrate that ATP can be UV cross-linked to the Runt domain and that ATP binding is sensitive to an amino acid substitution in the putative Kinase-1a motif (P-loop).

Adenosine Triphosphate↗
PubMed1996

Cloning and characterization of cDNA for adenosine kinase from mammalian (Chinese hamster, mouse, human and rat) species. High frequency mutants of Chinese hamster ovary cells involve structural alterations in the gene.

The enzyme adenosine kinase constitutes the major purine nucleoside phosphorylating activity in mammalian cells. In view of its central role in adenosine metabolism, which is an important physiological regulator, an understanding of the primary structure of adenosine kinase is of much interest. Using microsequence information from peptides derived from purified Syrian hamster liver enzyme, we have succeeded in isolating full length cDNA clones encoding adenosine kinase from Chinese hamster ovary cells and mouse 3T3 cells. The open reading frames in these clones consist of 334 and 335 amino acids and encode proteins of molecular masses 37364 Da and 37489 Da, respectively. In addition, the coding and upstream sequences for adenosine kinase from human (HeLa cells) and rat liver have also been cloned and sequenced. Transfection of an adenosine-kinase-deficient mutant (selected for resistance to the adenosine analog toyocamycin) of Chinese hamster ovary cells with a plasmid containing the cloned adenosine kinase cDNA, leads to regaining of adenosine kinase activity in the transformed cell. The adenosine kinase transformants also simultaneously lost their toyocamycin resistance and became similarly sensitive to the analog as the parental wild-type Chinese hamster ovary cells. The cloned adenosine kinase cDNA was also used to examine structural changes in mutants affected in adenosine kinase. In Chinese hamster ovary cells, one type of mutant that lacks adenosine kinase activity and displays high degree of resistance to various adenosine analogs, is obtained at an unusually high spontaneous frequency (10(-4)-10(-3)). Results of Southern and northern-blot analysis provide evidence that this group of mutants involves gross structural alterations affecting the adenosine kinase gene. Such structural alterations are not observed in another type of mutant which exhibits increased resistance only to C-adenosine analogs. Sequence similarity searches indicate that several of the bacterial and yeast sugar kinases (ribokinase, fructokinase and inosine-guanosine kinase) exhibit limited but significant similarity to the mammalian adenosine kinase. The sequence similarity data support the possibility that adenosine kinase shares a common evolutionary ancestor with these protein sequences.

Adenosine Kinase↗
PubMed1996

Neutralization of SIVmac239/17E in lymphocyte cultures involves virus strain-specific linear and conformational epitopes encoded by different regions of the env gene including the "V3" domain.

SIVmac251 and its closely related derivatives SIVmac239 and SIVmac239/17E vary greatly in their susceptibility to neutralization with homologous and heterologous antisera. Whereas SIVmac251 induces homologous neutralizing antibodies, the antibodies induced by SIVmac239 rarely neutralize infectivity of this virus in lymphocyte cultures. In contrast, SIVmac239/17E is remarkably susceptible to neutralization with homologous and heterologous antisera induced by other strains of SIVmac. In this study, we studied the molecular basis for the neutralization of SIVmac239/17E. Using chimeric viruses in which different regions of the env gene of both SIVmac239 and SIVmac239/17E were inserted into a background of either of the parental genomes, we showed that the newly acquired neutralization properties of SIVmac239/17E were attributable to amino acid substitutions between the V2 and V4 regions of gp 120. Site-directed mutagenesis of the env gene in this region showed that the arginine substitutions at positions 334 and/or 340 within the "V3" domain were fundamental to virus neutralization but other substitutions in the V2-V4 region added to the ease of its neutralization since it became neutralizable with much higher dilutions of serum. The molecular determinants for neutralization of this virus are distinct from those reported as responsible for neutralization of SIVmac251 and both are distinct from SIVmac239.

Animals↗
PubMed1996

Assignment of free and disulfide-bonded cysteine residues in testis angiotensin-converting enzyme: functional implications.

Human testicular angiotensin-converting enzyme (tACE) is an extracellular protein that contains seven cysteine residues. The cysteines occur in a sequential distribution that is precisely mimicked in the tACE from rabbit and mouse, and in both domains of all known species of somatic ACE. One of the cysteines in human tACE, Cys496, is present in the reduced form as shown by labeling it with 5-[[2-(iodoacetyl)amino]ethylamino]naphthalene-1-sulfonic acid, isolating the fluorescent peptide from enzymatic digests by HPLC, and analyzing its sequence by matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS). Thiol reagents have no significant effect on the activity of tACE, indicating that this Cys is not involved in catalysis. The other six cysteines exist as three disulfides. Mass spectral analysis of cyanogen bromide peptides has established that the cystine connectivities follow a nearest-neighbor, aabbcc, pattern i.e., Cys152-Cys158, Cys352-Cys370, and Cys538-Cys550, in which the disulfides form three small loops of five, 17, and 11 residues, respectively. Although these disulfide loops constitute less than 5% of the total sequence of the protein, they contribute to the overall structural stabilization of tACE.

Amino Acid Sequence↗
PubMed1996

A point mutation in interleukin-2 that alters ligand internalization.

In previous studies, we have identified an interleukin-2 (IL-2) analog containing a point mutation at position 51 (T51P) that expresses nearly wild-type bioactivity, yet has approximately 10-fold lower receptor binding affinity. Since ligand-dependent receptor internalization may be the rate-limiting step controlling the duration of IL-2 receptor signaling, a reduction in the receptor internalization rate could contribute to the observed response enhancement for this analog. To evaluate this possibility, we compared the internalization of IL-2 and T51P in three separate assays. While the internalization rate for IL-2 agreed with values determined by others, the internalization of T51P was markedly reduced. The receptor binding rate constants for this analog were only slightly different; thus, altered binding kinetics could not explain the decreased internalization rate. The effects of reduced internalization were also observable in bioassays, where T51P maintained T-cell proliferation for a longer period compared with IL-2. These results indicate that the T51P point mutation reduces the receptor internalization rate compared with IL-2 in a fashion that is independent of the dissociation rate. This analog may represent a new approach to the preparation of cytokine analogs with potentiated agonist and antagonist properties.

Binding Sites↗
PubMed1996

Interaction of nitric oxide synthase with the postsynaptic density protein PSD-95 and alpha1-syntrophin mediated by PDZ domains.

Neuronal nitric oxide synthase (nNOS) is concentrated at synaptic junctions in brain and motor endplates in skeletal muscle. Here, we show that the N-terminus of nNOS, which contains a PDZ protein motif, interacts with similar motifs in postsynaptic density-95 protein (PSD-95) and a related novel protein, PSD-93.nNOS and PSD-95 are coexpressed in numerous neuronal populations, and a PSD-95/nNOS complex occurs in cerebellum. PDZ domain interactions also mediate binding of nNOS to skeletal muscle syntrophin, a dystrophin-associated protein. nNOS isoforms lacking a PDZ domain, identified in nNOSdelta/delta mutant mice, do not associate with PSD-95 in brain or with skeletal muscle sarcolemma. Interaction of PDZ-containing domains therefore mediates synaptic association of nNOS and may play a more general role in formation of macromolecular signaling complexes.

Amino Acid Sequence↗
PubMed1996

A binding site for chlorambucil on metallothionein.

It is of interest to test the hypothesis that induced metallothionein (MT) acts in acquired drug resistance by covalent sequestration. In this study MT was incubated in vitro with chlorambucil (CHB) under conditions where only 1:1 covalent adducts were formed. The proteolytic products of these adducts were analyzed by HPLC and mass spectrometry to reveal two major sites of modification. These were the sulfur atoms of cysteines 33 and 48, which cochelate the same metal atom in native MT. The time course of the reaction was followed using on-line electrospray ionization with a double-focusing mass spectrometer. These experiments showed that drug-modified MT binds seven metal ions, as does the unmodified protein. Molecular docking experiments showed that the selectively of drug binding is influenced by the presence of the aziridinium ion in the drug structure and complementary charge densities in the protein structure.

Amino Acid Sequence↗
PubMed1996

Kinetic analysis of ligand binding to interleukin-2 receptor complexes created on an optical biosensor surface.

The interleukin-2 receptor (IL-2R) is composed of at least three cell surface subunits, IL-2R alpha, IL-2R beta, and IL-2R gamma c. On activated T-cells, the alpha- and beta-subunits exist as a preformed heterodimer that simultaneously captures the IL-2 ligand as the initial event in formation of the signaling complex. We used BIAcore to compare the binding of IL-2 to biosensor surfaces containing either the alpha-subunit, the beta-subunit, or both subunits together. The receptor ectodomains were immobilized in an oriented fashion on the dextran matrix through unique solvent-exposed thiols. Equilibrium analysis of the binding data established IL-2 dissociation constants for the individual alpha- and beta-subunits of 37 and 480 nM, respectively. Surfaces with both subunits immobilized, however, contained a receptor site of much higher affinity, suggesting the ligand was bound in a ternary complex with the alpha- and beta-subunits, similar to that reported for the pseudo-high-affinity receptor on cells. Because the binding responses had the additional complexity of being mass transport limited, obtaining accurate estimates for the kinetic rate constants required global fitting of the data sets from multiple surface densities of the receptors. A detailed kinetic analysis indicated that the higher-affinity binding sites detected on surfaces containing both alpha- and beta-subunits resulted from capture of IL-2 by a preformed complex of these subunits. Therefore, the biosensor analysis closely mimicked the recognition properties reported for these subunits on the cell surface, providing a convenient and powerful tool to assess the structure-function relationships of this and other multiple subunit receptor systems.

Animals↗
PubMed1996

Vibrio cholerae hemagglutinin/protease (HA/protease) causes morphological changes in cultured epithelial cells and perturbs their paracellular barrier function.

In this report, we describe the cytotoxic activity of the cholera hemagglutinin/protease (HA/protease). A concentrated protein sample from the 37 degrees C overnight culture supernatant of CVD110, a delta ctxA, delta zot, delta Ace and hlyA::(ctxB mer) mutant of El Tor biotype Ogawa serotype strain E7946 caused morphological changes in cultured MDCK-I epithelial cells and altered their arrangement of filamentous actin (F-actin) and Zonula occludens-associated protein ZO-1. The drastic morphological changes can be inhibited by Zincov, a specific bacterial metalloprotease inhibitor. The cytotoxic fractions of the sample after FPLC gelfiltration fractionation showed two visible protein bands with molecular weights of approximately 34- and 32 kDa. Microsequencing of these two proteins revealed that they were the cholera HA/protease.

Actins↗
PubMed1996

[Study of an ancient corpse of the Warring States period unearthed from Tomb No. 1 at Guo-Jia Gang in Jingmen City (A comprehensive study)].

An ancient female corpse excavated from the robbed tomb No. 1 at Guo-Jia-Gang in Jingmen city of Hubei Province was studied. It was confirmed that the corpse was buried more than 2,300 years ago, namely at the middle stage of the Warring States period and is the earliest one of the ancient coffin corpses unearthed in China so far. It is of the same type as the ancient corpses of Weatern Han Dynasty excavated from tomb No. 1 at Ma-Wang-Dui in Changsha City and from the tomb No. 168 at Phoenix Hill in Jinagling City respectively. The collagenous fibers of connective tissue from all parts in the corpse were well preserved, showing fine ultrastructure with clear periodical cross banding, and a lot of bacterial spores were found between them. In addition, a large number of eggs of clonorchis sinensis and trichuris trichiura were found in the content of the bowel. The level and the cause of good preservation were discussed and it was believed that the good preservation may result from the combined effect of the internal and external environmental factors around the corpse.

Animals↗
PubMed1996

The relationship between mast cell infiltration and the expression of PCNA and EGFr in experimental hepatocellular carcinoma of liver and squamous cell carcinoma of stomach in rats.

The expression of PCNA and EGFr in chemically induced hepatocellular carcinoma of liver and squamous cell carcinoma of stomach in rats were immunohistochemically observed. The results showed that the carcinoma cells of both tumors revealed a positive immunreaction to PCNA and EGFr. The histochemical observation of mast cell (MC) in both tumor tissues showed that the amount of MC in the surroundings of tumor cell nests was markedly different. According to the amount of the surrounding MCs the tumor cell nests could be divided into two groups: Group A with abundant MC infiltration and Group B with only scarce or without MC infiltration. The PCNA-positive cells in the tumor cell nests of both groups were calculated respectively. The results revealed that the amount of PCNA-positive cells in the group B was markedly more than that in the group A. The numerical ratio between two groups was 3:1 in the liver carcinoma and 2:1 in the stomach carcinoma approximately. An overexpression of EGFr was observed in tumor tissues of both groups, but there was also a marked difference in the amount of positively expressed cells and in the intensity of their staining reaction between both groups. The positively expressed cells in group B were much more and their staining intensity was much stronger than those in group A. According to the above mentioned results of observation, the expression state of both factors (PCNA and EGFr) was basically identical, suggesting that the MC may possess some inhibitory effect upon the growth rate of tumor cells of the experimental hepatocellular carcinoma of liver and the squamous cell carcinoma of stomach in rats.

Animals↗
PubMed1996

Persistently seronegative men from whom HIV-1 has been isolated are genetically and immunologically distinct.

Studies in both monkeys and humans have suggested that transient infection with HIV-1 can occur without provoking a measurable humoral immune response. The objective of this study was to look for genetic and immunologic correlates of transient HIV-1 infection in antibody-negative men from whom HIV-1 had been isolated. The distributions of MHC class I, class II, and TAP (transporter protein associated with antigen processing) region genes were compared between 23 persistently seronegative men from whom HIV-1 was isolated at least once (isol+/Ab-) and 137 men who seroconverted. A subset of 13 of the 23 isol+/Ab- men were compared to 27 seronegative men for distribution of CD25+CD4+ and CD25+CD8+ cells in the absence of exogenous immunologic stimulation. The prevalences of the TAP1.4, and a combination of TAP1.4, and TAP2.3 variants were significantly higher in the isol+/Ab- men. The proportion of CD8+ cells that expressed CD25+ antigen was also significantly higher in the isol+/Ab- men than in the seronegative men. We conclude that isol+/Ab- men may be genetically and immunologically distinct from HIV-1 susceptible men. We hypothesize that activated CD8+ cells may have cleared HIV-1 infection in these men through genetically mediated influences of the TAP genes on the presentation of peptides by HLA class I molecules.

ATP Binding Cassette Transporter, Subfamily B, Mem↗