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Biomedical subjects

J Wu

Publications and source records attributed to J Wu.

At least 1,261 records · Page 70Linked to original sources

Ultrasound heating in a tissue-bone phantom.

Temperature rise generated by focused ultrasound beams was tested on semipermanent tissue-bone phantoms. The phantoms were capped (sealed) plastic hollow cylindrical containers filled with tissue-mimicking material (TMM), in which were imbedded 25 microns diameter copper-constantan thermocouples (TC) and a piece of compact human or cow bone. The acoustic frequency specific attenuation coefficient of TMM was adjusted to be 0.3 dB cm-1 MHz-1 as specified by the FDA for a frequency range of 1-5 MHz. A high density 0.318 cm thick polyethylene sheet was chosen as the material to make caps of the phantoms. A formula developed to estimate the upper limit of temperature rises at tissue-bone interfaces generated by focused ultrasound has been proved to be appropriate experimentally using the semipermanent phantoms.

Animals↗

Fabrication and characterization of glucose sensors based on a microarray H2O2 electrode.

A glucose sensor was recently developed by modifying an immobilized GOD (glucose oxidase) membrane and coating Nafion diffusion limiting membrane over an amperometric H2O2 MAE (microarray electrode), which was fabricated by using standard planar processing. The chip of the sensor is 2.5 mm by 15. The MAE is composed of an Ag/AgCl RE (reference electrode), a Pt CE (counter electrode) and two sets of Pt WE (working electrode), each set consists of five bands. Silicon nitride was chosen as the top insulator. Six MAEs different in both width and length of the band were examined and the MAE with the narrowest width (10 microns) of bands exhibits the best electrochemical performances. The glucose sensor based on the MAE B1-2 presents a linear range of 0.5-40 mM glucose at 37 degrees C with a 1/15 M phosphate buffer solution (pH = 7.0) as supporting electrolyte. The sensitivity, background current, response time, CV (coefficient of variation) and long-term stability of the sensor are 7.1 +/- 0.5 nA/mM, 2 +/- 0.5 nA, 30 s, 3.4% and 6 days respectively.

Biosensing Techniques↗

Insulin secretion in pancreatic islets from rats with cirrhosis.

Cirrhosis was induced in rats by subcutaneous injections of CCl4 for 13 or 17 weeks. The morphology of the pancreatic islets from the CCl4-treated rats was found to be normal. The CCl4-treated rats had lower fasting serum glucose levels and higher serum insulin levels than the controls. After an oral glucose load (3 g/kg body weight), glucose levels in CCl4-treated rats stayed within the normal range, whereas the serum insulin levels remained higher with a delayed decline of insulin with time. In vitro perifusion of islets from the CCl4-treated rats showed that the response to 16.7 mmol/l glucose was reduced with both lower total insulin output and stimulated insulin output, whereas the patterns of first and second phase of insulin release did not differ. The insulin content of the perifused islets was not affected by 13 weeks of CCl4 treatment. Islets from rats treated with CCl4 for 17 weeks showed normal secretory response to 20 mmol/l L-arginine. Taken together, the results, showing normal or reduced capacity for insulin secretion, suggest that the hyperinsulinemia accompanying CCl4-induced cirrhosis is not due to increased secretion of the pancreatic islets. It may rather be associated with decreased insulin degradation by the liver with cirrhosis.

Animals↗

Patterns of cortical activity in schizophrenia.

Eighty-three patients with schizophrenia and 47 healthy controls received positron emission tomography (PET) with 18F-2-deoxyglucose uptake while they were executing the Continuous Performance Test (CPT). The entire cortex was divided into 16 regions of interest in each hemisphere, four in each lobe of the brain, and data from corresponding right and left hemispheric regions were averaged. Data from the schizophrenic patients were subjected to a factor analysis, which revealed five factors that explained 80% of the common variance. According to their content, the factors were identified and labelled 'parietal cortex and motor strip', 'associative areas', 'temporal cortex', 'hypofrontality' (which included midfrontal and occipital areas) and 'frontal cortex'. Hemispheric asymmetry was only confirmed for the temporal cortex. Factor weights obtained in the schizophrenic group were applied to the metabolic data of the healthy controls and factors scales computed. Schizophrenics were significantly more hypofrontal than the controls, with higher values on the 'parietal cortex and motor strip' factor and a trend towards higher values in the temporal cortex. A canonical discriminant analysis confirmed that the 'hypofrontality' and 'parietal cortex and motor strip' factors accurately separated the schizophrenic group from the healthy controls. Hemispheric asymmetry was only confirmed for the temporal lobe. Significantly higher factor scores for the left temporal lobe in schizophrenics than in normals were obtained when calculated for the right and left hemisphere separately. Taken together, our results confirm the importance of hypofrontality as a pattern of cortical metabolic rate and point to the potential importance of parietal and motor strip function in schizophrenia.

Adult↗

The murine Xe169 gene escapes X-inactivation like its human homologue.

Among a number of genes that escape X-chromosome inactivation in humans, three have been evaluated in mice and unexpectedly all three are subject to X-inactivation. We report here the cloning and expression studies of a novel mouse gene, Xe169, and show that it escapes X-inactivation like its human homologue. Xe169 was assigned to band F2/F3 on the mouse X chromosome by fluorescent in situ hybridization and Southern analysis indicates that the gene is located outside the pseudoautosomal region. Homologous, but divergent, sequences exist on the Y chromosome. In vitro and in vivo studies show that Xe169 is expressed from both the active and the inactive X chromosomes. Xe169 is the first cloned non-pseudoautosomal gene that escapes X-inactivation in mice.

Animals↗

The LEC rat has a deletion in the copper transporting ATPase gene homologous to the Wilson disease gene.

The Long-Evans Cinnamon (LEC) rat shows similarity to Wilson disease in many clinical and biochemical features. We have cloned cDNAs for the rat gene (Atp7b) homologous to the human Wilson disease gene (ATP7B) and have used them to identify a partial deletion in the Atp7b gene in the LEC rat. The deletion removes at least 900 bp of the coding region at the 3' end, includes the crucial ATP binding domain and extends downstream of the gene. Our results provide convincing evidence for defining the LEC rat as an animal model for Wilson disease. This model will be important for studying liver pathophysiology, for developing therapy for Wilson disease and for studying the pathway of copper transport and its possible interaction with other heavy metals.

Adenosine Triphosphatases↗

A 300 kilobase interval genetic map of rice including 883 expressed sequences.

We have constructed a high resolution rice genetic map containing 1,383 DNA markers at an average interval of 300 kilobases (kb). The markers, distributed along 1,575 cM on 12 linkage groups, comprise 883 cDNAs, 265 genomic DNAs, 147 randomly amplified polymorphic DNAs (RAPD) and 88 other DNAs. cDNAs were derived from rice root and callus, analysed by single-run sequencing and searched for similarities with known proteins. Nearly 260 rice genes are newly identified and mapped, and genomic DNA and cloned RAPD fragments were also sequenced to generate STSs. Our map is the first significant gene expression map in plants. It is also the densest genetic map available in plants and the first to be backed up comprehensively by clone sequence data.

Chromosome Mapping↗

Isolation and characterization of XE169, a novel human gene that escapes X-inactivation.

Overlapping cDNA clones for a novel human X-linked gene, XE169, have been isolated and characterized. The composite cDNA sequence comprises 5910 bp (or 5901 bp) plus a poly(A) tail, with a 531 bp 5' and 696 bp 3' untranslated regions. The sequence represents a full-length or near full-length cDNA for the gene since Northern blot analysis reveals only a single prominent band approximately 6 kb in size. Alternative splicing generates two distinct transcripts either containing or missing a stretch of nine nucleotides in the XE169 single large open reading frame, which in turn predict two XE169 protein isoforms composed of 1557 and 1560 amino acids, respectively. Southern hybridization analysis of a panel of human-mouse somatic cell hybrids containing various portions of translocated human X chromosomes has assigned XE169 to the proximal half of the X short arm between Xp21.1 and the centromere. XE169 is expressed in multiple human tissues tested and homologous sequences exist on the human Y chromosome and in the genomes of five other eutherian mammals examined. RT-PCR analysis of somatic cell hybrids containing either an active or an inactive human X chromosome on a rodent background demonstrated that XE169 escapes X-inactivation.

Alternative Splicing↗

Expression of a G-protein beta subunit-related gene during lymphocyte activation.

Using a subtractive strategy, we have cloned an activation-related gene from a human B cell cDNA library. Sequence analysis revealed that this gene was identical to H12.3, a gene belonging to an expanding family of guanine nucleotide-binding protein beta subunits. The expression of H12.3 was inducible in the late phase of mitogen-stimulated T and B cells. In T cells, IL-2 and IL-4 by themselves had no direct effect on the expression of H12.3, but they could augment the level of steady-state H12.3 mRNA stimulated by phytohemagglutinin. On the other hand, IFN-gamma and IL-6 had no obvious effect on the expression in B cells with or without Staphylococcus aureus Cowan I-stimulation. Cyclosporin A, a strong immunosuppressant, inhibited the mitogen-stimulated expression of H12.3, but rapamycin, another such agent, did not. In synchronized Jurkat cells, the expression of H12.3 had no cell cycle-dependent decrease in S and G2/M phase, while cyclin E, which controls the progression of the cell cycle in late G1 phase, did show a periodic expression pattern. The results suggest that H12.3 might be involved in regulation of lymphocyte activation.

Animals↗

Regulation of the Escherichia coli lrp gene.

Lrp (leucine-responsive regulatory protein) is a major Escherichia coli regulatory protein which regulates expression of a number of operons, some negatively and some positively. This work relates to a characterization of lrp, the gene encoding Lrp. Nucleotide sequencing established that the coding regions of lrp and trxB (encoding thioredoxin reductase) are separated by 543 bp and that the two genes are transcribed in opposite directions. In addition, we used primer extension, deletion analyses, and lrp-lacZ transcriptional fusions to delineate the promoter and regulatory region of the lrp operon. The lrp promoter is located 267 nucleotides upstream of the translational start codon of the lrp gene. In comparison with a wild-type strain, expression of the lrp operon was increased about 3-fold in a strain lacking Lrp and decreased about 10-fold in a strain overproducing Lrp. As observed from DNA mobility shift and DNase I footprinting analyses, Lrp binds to one or more sites within the region -80 to -32 relative to the start point of lrp transcription. A mutational analysis indicated that this same region is at least partly required for repression of lrp expression in vivo. These results demonstrate that autogenous regulation of lrp involves Lrp acting directly to cause repression of lrp transcription.

Bacterial Proteins↗

Identification of a new catenin: the tyrosine kinase substrate p120cas associates with E-cadherin complexes.

p120cas is a tyrosine kinase substrate implicated in ligand-induced receptor signaling through the epidermal growth factor, platelet-derived growth factor, and colony-stimulating factor receptors and in cell transformation by Src. Here we report that p120 associates with a complex containing E-cadherin, alpha-catenin, beta-catenin, and plakoglobin. Furthermore, p120 precisely colocalizes with E-cadherin and catenins in vivo in both normal and Src-transformed MDCK cells. Unlike beta-catenin and plakoglobin, p120 has at least four isoforms which are differentially expressed in a variety of cell types, suggesting novel means of modulating cadherin activities in cells. In Src-transformed MDCK cells, p120, beta-catenin, and plakoglobin were heavily phosphorylated on tyrosine, but the physical associations between these proteins were not disrupted. Association of p120 with the cadherin machinery indicates that both Src and receptor tyrosine kinases cross talk with proteins important for cadherin-mediated cell adhesion. These results also strongly suggest a role for p120 in cell adhesion.

3T3 Cells↗

Evaluation of the Humphrey FASTPAC threshold program in glaucoma.

The makers of the Humphrey perimeter have developed a rapid thresholding program, FASTPAC, to address the problems of fatigue and poor test performance associated with prolonged perimetry. The performance of FASTPAC was compared with the standard threshold program of the Humphrey visual field analyser (program 24-2) in 44 glaucoma patients. FASTPAC reduced the mean test time by 35% (from 12.6 to 8.2 minutes), owing to a reduction in the number of stimulus presentations. FASTPAC underestimated the mean deviation (MD) (Wilcoxon, p = 0.007) and corrected pattern standard deviation (CPSD) (Wilcoxon, p = 0.005). The sum of the differences between FASTPAC and the standard program was -1.19 (SD 2.37) dB for MD and 0.97 (2.14) dB for CPSD. The measured difference between the two methods was independent of the value of either MD or CPSD, indicating that the measurement error was just as likely to occur with either early or advanced visual field loss. No difference was noted in short term fluctuation or in the reliability indices of test performance. These results indicate that FASTPAC, while considerably faster, is not as accurate as the standard threshold program at measuring retinal sensitivity in glaucoma. This inaccuracy may be offset in practical terms by greater patient acceptability and by increasing the number of patients capable of performing reliable threshold perimetry because of the reduction in duration of the FASTPAC program.

Aged↗

Handwritten digit recognition using two-layer self-organizing maps.

In this paper, we present a two-layer self-organizing neural network based method for handwritten digit recognition. The network consists of a base layer self-organizing map and a set of corresponding maps in the second layer. The input patterns are partitioned into subspace in the first layer. Patterns in a subspace are led to the second layer and a corresponding map is built according to the first layer performance. In the classification process, each pattern searches for several closest nodes from the base map and then it is classified into a specified class by determining the nearest model of the corresponding maps in the second layer. The new method yielded higher accuracy and faster performance than the ordinary self-organizing neural network.

Handwriting↗

Palmitoyl carnitine modifies sodium currents and induces transient inward current in ventricular myocytes.

Long-chain acylcarnitines increase within 2 min in ischemic myocardium in vivo and induce delayed afterdepolarizations (DADs) and complex oscillations of membrane potential in vitro. This study was performed to assess the ionic currents underlying these electrophysiological alterations in isolated rabbit ventricular cells using whole cell voltage-clamp procedures. Palmitoyl carnitine (10 microM, for 6-10 min) elicited a transient inward current (Iti) in the presence of blockade of Ca2+ and K+ channels. The effect of palmitoyl carnitine was reversible after washout (n = 6). The amplitude of Iti was dependent on the amplitude of the preceding depolarization step. Palmitoyl carnitine (10 microM, for > 2 min) also induced another inward current, which was activated spontaneously at potentials between -120 and -20 mV with a linear current-voltage relationship (1.0 +/- 0.1 nA at -80 mV). This current was abolished by replacing extracellular Na+ with tetraethylammonium chloride, indicating that Na+ was the charge carrier. Inactivation of this current was slow (gamma = 885.9 +/- 89.1 ms, n = 12) or incomplete, indicating the appearance of a slow-inactivating Na+ inward current [INa(s)]. Palmitoyl carnitine always induced INa(s) before the appearance of Iti. Intracellular ethylene glycol-bis(beta-amino-ethyl ether)-N,N,N',N'-tetraacetic acid (10 mM) abolished Iti but did not suppress INa(s) (n = 4), indicating that INa(s) was not activated by intracellular Ca2+ (Cai2+). Tetrodotoxin (10 microM) also decreased the amplitude of INa(s). Thus palmitoyl carnitine induces INa(s), which likely leads to an increase in Na+ influx, thereby eliciting an increase in Cai2+ via the Na(+)-Ca2+ exchanger and leading to the development of Iti, DADs, and triggered activity.

Animals↗

Glucocorticoid regulation of an insulin-like growth factor-binding protein-4 protease produced by a rat neuronal cell line.

Insulin-like growth factor-binding protein-4 (IGFBP-4) is expressed in distinct regions in the rodent brain from the perinatal period into adulthood and is postulated to modulate the action of the insulin-like growth factors (IGFs) in vivo. This study was initiated to examine the regulation of IGF-binding protein-4 (IGFBP-4) in B104 cells, a rat neuronal cell line in which IGFBP-4 is the predominant secreted IGFBP. Exposure of B104 monolayer cultures to dexamethasone reduced native IGFBP-4 abundance to less than 10% of that in control medium by 48 h. Immunoblots showed that the decline in intact 24-kilodalton IGFBP-4 was accompanied by an increase in a 16-kilodalton immunoreactive fragment. In addition, IGFBP-4 proteolytic activity in medium was increased after exposure of the cells to dexamethasone. The protease was calcium dependent and appeared to be of the serine protease class, because activity could be inhibited by phenylmethylsulfonylfluoride and aprotinin, but not antipain, leupeptin, or pepstatin. Although the proteolytically modified IGFBP-4 retained the ability to bind IGFs, the affinities were approximately 13- and 20-fold lower for IGF-I and IGF-II, respectively. These data indicate that B104 cells produce an IGFBP-4 protease that is regulated by glucocorticoids. The actions of this protease reduce the affinity of IGFBP-4 for the IGFs without abolishing binding. Because both the IGFs and glucocorticoids have important roles in brain development, it is possible that some glucocorticoid actions in the brain could be mediated by proteolysis of IGFBP-4, which, in turn, would alter IGF action.

Animals↗

The lactose permease meets Frankenstein.

The lactose permease (lac) of Escherichia coli is a paradigm for membrane transport proteins. Encoded by the lacY gene, the permease has been solubilized, purified to homogeneity, reconstituted into phospholipid vesicles and shown to catalyse the coupled translocation of beta-galactosides and H+ with a stoichiometry of unity. Circular dichroism and other spectroscopic approaches demonstrate that the purified permease is about 80% helical. Based on hydropathy analysis of the primary amino-acid sequence, a secondary structure has been proposed in which the protein has 12 hydrophobic domains in alpha-helical conformation that traverse the membrane in zigzag fashion connected by hydrophilic loops. A variety of other approaches are consistent with the model and demonstrate that both the N and C termini are on the inner surface of the membrane, and studies on an extensive series of lac permease/alkaline phosphatase fusion proteins provide exclusive support for the topological predictions of the 12-helix motif. This presentation concentrates on the use of site-directed fluorescence spectroscopy to study structure-function relationships in the permease.

Amino Acid Sequence↗

A comparison of preservation techniques in a vascularized rat laryngeal transplant model.

In earlier laryngeal transplantation studies by Takenouchi, et al., the longest ischemic interval tolerated was 45 minutes. A new animal model and enhanced preservative solutions made reassessment timely. This study evaluated two determinants of graft viability: 1. the duration of ischemia and 2. the composition of the preservative media. Three groups of viable transplants were assessed. Groups I and II were preserved with iced heparinized saline with respective ischemic intervals of 3 and 6 hours. Group III was preserved with the Wisconsin solution during a 20-hour ischemic interval. All animals were sacrificed at 24 hours. Representative sections of group I confirmed viability whereas group II exhibited both clinical and histologic evidence of irreversible vascular change. In contrast, most representative sections in group III had little demonstrable change. These data suggest that laryngeal allografts can endure prolonged ischemic intervals if properly maintained.

Adenosine↗