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Biomedical subjects

C Wu

Publications and source records attributed to C Wu.

At least 235 records · Page 13Linked to original sources
PubMed1999

Thiols and polyamines in the cytoprotective effect of taurine on carbon tetrachloride-induced hepatotoxicity.

The mechanism by which taurine (2-aminoethanesulfonic acid) protects hepatocytes injury induced by carbon tetrachloride (CCl4) is not fully understood. In a previous study, we reported that cellular polyamines play an important role in this mechanism. The relationship between cellular glutathione (GSH), protein-SH levels, and lactate dehydrogenase (LDH), with respect to the effect of polyamine on the cytoprotective ability of taurine in CCl4-induced toxicity in isolated rat hepatocytes, was examined. CCl4 induced a LDH release and decreased cellular thiols and polyamine levels. Treating with taurine reversed these depletions. The effect of CCl4 was also reversed by the addition of exogenous polyamines. Pretreating with alpha-difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase, which is a key enzyme in polyamine biosynthesis and therefore used to deplete cellular polyamine, prevented the protective effect of taurine. Adding diethyl maleate, a cellular glutathione-depleting agent, reduced the effect of exogenous polyamines. The role of polyamine in the cytoprotective effect of taurine in CCl4-induced toxicity may therefore be by preventing, among others, GSH and protein-SH depletions.

Animals↗
PubMed1999

Characterization of mouse interleukin-12 p40 homodimer binding to the interleukin-12 receptor subunits.

Interleukin-12 (IL-12) is a heterodimeric cytokine composed of two disulfide-bonded subunits, p35 and p40, which has important regulatory effects on T cells and natural killer (NK) cells. In contrast to heterodimeric IL-12, a homodimer of the p40 subunit, designated (p40)2, has been shown to be a potent IL-12 antagonist. To study the interaction between (p40)2 and the known IL-12 receptor (IL-12R) subunits, IL-12Rbeta1 and IL-12Rbeta2, we directly measured the binding activity of mouse (p40)2 to ConA-activated lymphoblasts and purified B cells from splenocytes of C57BL/6J mice. These results demonstrated the presence of both high (Kd about 5 pM) and low affinity (Kd about 15 nM) binding sites for mouse 125I-labeled (p40)2. To elucidate which of the IL-12R subunits binds mouse (p40)2, binding studies of mouse 125I-labeled (p40)2 to Ba/F3 cells expressing recombinant mouse IL-12Rbeta1 and/or mouse IL-12Rbeta2 were carried out. Mouse IL-12Rbeta1 bound mouse 125I-labeled (p40)2 with high and low affinities, comparable to that observed on Con A blasts and B cells. In contrast, mouse IL-12Rbeta2 bound mouse 125I-labeled (p40)2 very poorly. These data demonstrate that similar to IL-12, mouse (p40)2 binds with both high and low affinity to Con A blasts and B cells, and that IL-12Rbeta1 is responsible for mediating the specific binding of mouse (p40)2.

Animals↗
PubMed1999

Synthesis and relaxometry of high-generation (G = 5, 7, 9, and 10) PAMAM dendrimer-DOTA-gadolinium chelates.

A series of high-generation (G) ethylenediamine-core polyamidoamine (PAMAM) dendrimers corresponding to G = 5, 7, 9, and 10 were conjugated with the bifunctional chelate 2-(4-isothiocyanatobenzyl)- 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetate (p-SCN-Bz-DOTA). Gadolinium (III) ion was added to the macromolecules, and the 1/T1 and 1/T2 NMRD profiles were measured at 3 degrees, 23 degrees, and 37 degrees C. The synthesis resulted in preparations that ranged from an average of 127 chelates and 96 Gd3+ ions per G = 5 dendrimer to an average of 3727 chelates and 1860 Gd3+ ions per G = 10 dendrimer. At 20 MHz and 23 degrees C, the 1/T1 ion relaxivity increased from 30 mM(-1) s(-1) for the G = 5 to 35 mM(-1) s(-1) for the G = 7 PAMAM dendrimer-DOTA-Gd, reaching a plateau at 36 mM(-1) s(-1) for the G = 9 and G = 10 dendrimers. A similar plateau was observed for 1/T2 with values of 36 mM(-1) s(-1) for G = 5, 42 mM(-1) s(-1) for G = 7, and 45 mM(-1) s(-1) for the G = 9 and G = 10 dendrimers. This "saturation" of ion relaxivity for high-generation dendrimers occurred over the entire frequency range studied. The 1/T1 and 1/T2 relaxivities decreased as the temperature decreased for each generation of dendrimer studied, implying that slow water exchange of bound water molecules with the bulk solvent limits the relaxivity. In such circumstances, increases in the rotational correlation time of the macromolecules associated with higher generations of dendrimer does not result in significant increases in the ion relaxivity. Although the ion relaxivity does not increase, the total molecular relaxivities increased from 2880 mM(-1) s(-1) to 66960 mM(-1) s(-1) for the G = 5 to the G = 10 dendrimer. The current findings are relevant for the design of high-generation dendrimer-based receptor agents.

Contrast Media↗
PubMed1999

Activated astrocytes display increased 5-HT2a receptor expression in pathological states.

In human brain tissues from patients dying with cerebral infarction, hypertensive encephalopathy, Alzheimer's disease, Huntington's disease, frontotemporal dementia, and Creutzfeldt-Jakob disease there is an activation of astrocytes. Such activated astrocytes display GFAP and strong 5-HT(2A), but not 5-HT(2B) or 5-HT(2C), receptor immunoreactivity; this 5-HT(2A) reaction has not been observed in normal, nonactivated astrocytes. It is suggested that an up-regulation of 5-HT(2A) receptors may be part of an early response reaction in astrocytes, possibly designed to maintain homeostasis or to induce secondary message pathways involving trophic factors or glycogenolysis.

Aged↗
PubMed1999

Surgical treatment of neuroblastoma.

Nine patients with neuroblastoma were surgically treated and postoperatively followed up. Of the 9 cases, 8 survived well while the other one died of extensive metastasis within cranium 2 years after the operation. One patient had stayed alive for over 12 years.

Adolescent↗
PubMed1999

Experimental study on detached renal tubular epithelial cells in urine of nephropathia epidemic patients.

To elucidate the pathogenesis of acute renal failure (ARF) with nephropathia epidemic (NE), provide experimental evidence for the new therapy to NE and observe the effects of Arg-Gly-Asp (RGD) peptides on adhesion of renal tubular epithelial cell (RTEC), urine specimens of patients were collected under sterile conditions. Detached RTECs were separated, cultured and identified. Hantan Virus antigen was determined by using indirect immunofluorescence method and effects of RGD on adhesion of RTECs was observed by subgroup counting as well as by flow cytometry. This study showed that: (1) sublethal RTECs existed in the urine of NE-ARF patients, which could be cultured in monolayer form; (2) there was NE antigen in RTECs; and (3) adhesion of RTECs could be inhibited by RGD.

Acute Kidney Injury↗
PubMed1999

Ultrasound velocity of trabecular cubes reflects mainly bone density and elasticity.

Studies have indicated that quantitative ultrasound (QUS) variables may be influenced by the mechanical properties of bone which in turn are determined by bone's material and structural properties. However, from these studies it is unclear what role density, elasticity, and structure play in determining velocity. Eighteen defatted, 12-mm cubic trabecular bone specimens were cut from cadaveric specimens. Amplitude-dependent speed of sound (SOS) using a single point QUS system was assessed in three orthogonal axes. Magnetic resonance images were obtained, from which measures of apparent trabeuclar structure were derived. The specimens were nondestructively tested in compression along three orthogonal axes defined by the sides of the cubes. The elastic modulus (in the three directions) and the strength (in one direction) were determined. Trabecular BMD was measured by quantitative computed tomography. SOS varied significantly with direction of measurement, with the highest value in the axial direction (axial:1715 m/s, sagittal: 1662 m/second, and coronal: 1676 m/s). SOS of each of the three axes was generally associated with the various mechanical (r = 0.30-0.87), density (r = 0.81-0.93), and bone structural variables (0.3-0.8). However, after adjusting the SOS correlations by density, only the correlation with elasticity remained significant in the coronal direction. BMD alone explained 88-93% of variance in SOS whereas in the multivariate model, BMD plus elasticity and/or anisotropic variables explained 96-98% of the variance in SOS. Variability of SOS is explained mostly by density and to a small extent by elasticity or anisotropy. Since only 2-6% of the variance of the QUS measurement is not explained by density and elasticity, one could conclude that the remaining variance reflects other properties of bone or perhaps simply measurement error. Evidence that these other properties may be structure related is only found in the anisotropy of QUS parameter.

Adult↗
PubMed1999

Construction and characterization of bacterial artificial chromosome libraries from the silkworm, Bombyx mori.

Two bacterial artificial chromosome (BAC) libraries were constructed using nuclear DNA from posterior silkglands of the silkworm (Bombyx mori) strains p50 and C108. The libraries contain a total of 36,864 clones, or approximately 9 genome equivalents. The average insert sizes in the libraries were 134.5 kb and 120.8 kb, respectively. PCR-based screening was performed on the p50 library using probes for 34 sequence-tagged sites (STSs). Between 3 and 11 (6.1 hits on average) clones were isolated with each STS, in good agreement with the library size, 5.8 genome equivalents. The previously reported close linkage between the Bombyx homologs of the invected (Bm in) and engrailed (Bm en) genes was confirmed by construction of a BAC contig that contained both. Moreover, screening revealed novel information about the chromosomal organization of the sericin-1 and DH-PBAN genes, which were localized within a 22-kb interval and are divergently oriented. These results show that it is possible to construct contigs and analyze chromosome organization using these libraries.

Animals↗
PubMed1999

An in vitro investigation of the dependence on sample thickness of the speed of sound along the specimen.

To measure the speed of sound (SOS), most quantitative ultrasound (QUS) devices use the transmission mode, whereby two transducers are placed on opposite sides of the sample. This mode is limited to a few specific skeletal sites because of the varying configuration of bone geometry and varying amounts of overlying soft tissue at most other sites. The aim of this study was to address the dependence of SOS measured along the sample on the thickness and composition of the bone sample. Bovine samples from mid-femur and trochanter, and perspex phantoms were used. We prepared the perspex samples in the shapes of blocks and cylinders to investigate the effect of wall thickness on SOS. The thickness of the blocks was decreased in decrements of 1 mm; a 22 mm diameter hole was drilled through the cylindrical samples and the hole size was gradually increased. The second configuration was also used with the bovine samples. For each experimental set-up five SOS measurements were acquired, with the probe aligned along the sample and a mean value computed. All measurements were taken with castor oil as the coupling agent, and in the cylindrical cases, the oil was used to fill the tube. The measurement precision determined as the root mean square coefficient of variation (RMSCV) was determined to be 0.14% and 0.65% for perspex and bovine samples respectively. The measured SOS on the perspex phantom (2760+/-4 m/s) was within the published values for bulk velocity. It was observed that for both perspex and bovine samples the SOS was independent of sample wall thickness greater than the wavelength (2.2 mm, 2.7 mm and 3.5 mm for perspex, trochanter and mid-femur respectively). The SOS decreased with sample wall thickness smaller than the wavelength in concordance with theoretical predictions. The SOS values obtained for bovine samples reflected either totally cortical (mid-femur) or a composite of cortical and cancellous bone (trochanter).

Animals↗
PubMed1999

Synthesis of furanonaphthoquinones with hydroxyamino side chains.

Several furanonaphthoquinones have shown useful activity in a yeast assay for DNA-damaging agents and cytotoxicity in mammalian cell culture assays. These results, together with the planar aromatic character of the furanonaphthoquinones, suggested that they might be acting as DNA intercalators. In an attempt to improve this activity, various analogues containing a hydroxyamino side chain have been synthesized. The analogues were prepared by standard methods, but some unexpected reactions were observed nonetheless. Thus, 8-formyl-5-methoxy-4,9-dihydronaphtho[2,3-b]furan-4,9-dione (24) showed an unusual reactivity toward reductive amination, with the reaction proceeding further to give one of two different cyclized products, depending on the amination reagent used. Bioassay results indicated that only simple furanonaphthoquines showed activity in a yeast assay for DNA-damaging agents; compounds with a substituted hydroxyamino side chain were uniformly inactive in this assay. Most of the compounds with a substituted hydroxyamino side chain on the furan ring did, however, show cytotoxicity, although none of them was any more active than the simple aldehyde 2-formyl-4, 9-dihydronaphtho[2,3-b]furan-4,9-dione (14). This evidence tends to suggest that the furanonaphthoquinones do not serve primarily as DNA intercalators, because if this were the case, they would have been expected to show an increased activity on conversion to their hydroxyamino side chain derivatives.

Animals↗
PubMed1999

Spontaneous CSF rhinorrhoea secondary to a middle cranial fossa defect.

Spontaneous cerebrospinal fluid rhinorrhoea is a clinical entity that is difficult to both diagnose and treat. A case in a 56-year-old woman with a defect in the lateral wall of the sphenoid sinus and middle cranial fossa floor is presented. Pathogenesis and management of this rare condition are discussed. Copyright 1999 Harcourt Publishers Ltd.

Journal Article↗
PubMed1999

Functional characterization of the interaction of Ste50p with Ste11p MAPKKK in Saccharomyces cerevisiae.

The Saccharomyces cerevisiae Ste11p protein kinase is a homologue of mammalian MAPK/extracellular signal-regulated protein kinase kinase kinases (MAPKKKs or MEKKs) as well as the Schizosaccharomyces pombe Byr2p kinase. Ste11p functions in several signaling pathways, including those for mating pheromone response and osmotic stress response. The Ste11p kinase has an N-terminal domain that interacts with other signaling molecules to regulate Ste11p function and direct its activity in these pathways. One of the Ste11p regulators is Ste50p, and Ste11p and Ste50p associate through their respective N-terminal domains. This interaction relieves a negative activity of the Ste11p N terminus, and removal of this negative function is required for Ste11p function in the high-osmolarity glycerol (HOG) pathway. The Ste50p/Ste11p interaction is also important (but not essential) for Ste11p function in the mating pathway; in this pathway binding of the Ste11p N terminus with both Ste50p and Ste5p is required, with the Ste5p association playing the major role in Ste11p function. In vitro, Ste50p disrupts an association between the catalytic C terminus and the regulatory N terminus of Ste11p. In addition, Ste50p appears to modulate Ste11p autophosphorylation and is itself a substrate of the Ste11p kinase. Therefore, both in vivo and in vitro data support a role for Ste50p in the regulation of Ste11p activity.

Adaptor Proteins, Signal Transducing↗
PubMed1999

An analysis of transvection at the yellow locus of Drosophila melanogaster.

Studies of a wide variety of organisms have shown that homologous sequences can exert a significant impact on each other, resulting in changes in gene sequence, gene expression, chromatin structure, and global chromosome architecture. Our work has focused on transvection, a process that can cause genes to be sensitive to the proximity of a homologue. Transvection is seen at the yellow gene of Drosophila, where it mediates numerous cases of intragenic complementation. In this article, we describe two approaches that have characterized the process of transvection at yellow. The first entailed a screen for mutations that support intragenic complementation at yellow. The second involved the analysis of 53 yellow alleles, obtained from a variety of sources, with respect to complementation, molecular structure, and transcriptional competence. Our data suggest two ways in which transvection may be regulated at yellow: (1) a transcriptional mechanism, whereby the ability of an allele to support transvection is influenced by its transcriptional competency, and (2) a structural mechanism, whereby the pairing of structurally dissimilar homologues results in conformational changes that affect gene expression.

Alleles↗
PubMed1999

Serodiagnosis of human granulocytic ehrlichiosis by a recombinant HGE-44-based enzyme-linked immunosorbent assay.

Current antibody testing for human granulocytic ehrlichiosis relies predominantly on indirect fluorescent-antibody assays and immunoblot analysis. Shortcomings of these techniques include high cost and variability of test results associated with the use of different strains of antigens derived from either horses or cultured HL-60 cells. We used recombinant protein HGE-44, expressed and purified as a maltose-binding protein (MBP) fusion peptide, as an antigen in a polyvalent enzyme-linked immunosorbent assay (ELISA). Fifty-five normal serum samples from healthy humans served as a reference to establish cutoff levels. Thirty-three of 38 HGE patient serum samples (87%), previously confirmed by positive whole-cell immunoblotting, reacted positively in the recombinant ELISA. In specificity analyses, serum samples from patients with Lyme disease, syphilis, rheumatoid arthritis, and human monocytic ehrlichiosis (HME) did not react with HGE-44-MBP antigen, except for one sample (specificity, 98%). We conclude that recombinant HGE-44 antigen is a suitable antigen in an ELISA for the laboratory diagnosis and epidemiological study of HGE.

Antibodies, Bacterial↗
PubMed1999

The LIM-only protein PINCH directly interacts with integrin-linked kinase and is recruited to integrin-rich sites in spreading cells.

PINCH is a widely expressed and evolutionarily conserved protein comprising primarily five LIM domains, which are cysteine-rich consensus sequences implicated in mediating protein-protein interactions. We report here that PINCH is a binding protein for integrin-linked kinase (ILK), an intracellular serine/threonine protein kinase that plays important roles in the cell adhesion, growth factor, and Wnt signaling pathways. The interaction between ILK and PINCH has been consistently observed under a variety of experimental conditions. They have interacted in yeast two-hybrid assays, in solution, and in solid-phase-based binding assays. Furthermore, ILK, but not vinculin or focal adhesion kinase, has been coisolated with PINCH from mammalian cells by immunoaffinity chromatography, indicating that PINCH and ILK associate with each other in vivo. The PINCH-ILK interaction is mediated by the N-terminal-most LIM domain (LIM1, residues 1 to 70) of PINCH and multiple ankyrin (ANK) repeats located within the N-terminal domain (residues 1 to 163) of ILK. Additionally, biochemical studies indicate that ILK, through the interaction with PINCH, is capable of forming a ternary complex with Nck-2, an SH2/SH3-containing adapter protein implicated in growth factor receptor kinase and small GTPase signaling pathways. Finally, we have found that PINCH is concentrated in peripheral ruffles of cells spreading on fibronectin and have detected clusters of PINCH that are colocalized with the alpha5beta1 integrins. These results demonstrate a specific protein recognition mechanism utilizing a specific LIM domain and multiple ANK repeats and suggest that PINCH functions as an adapter protein connecting ILK and the integrins with components of growth factor receptor kinase and small GTPase signaling pathways.

Adaptor Proteins, Signal Transducing↗