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Biomedical subjects

C H Chu

Publications and source records attributed to C H Chu.

At least 37 records · Page 2Linked to original sources
PubMed1999

Detection and identification of Mycobacterium tuberculosis by nested PCR assays in cerebrospinal fluid samples from patients with suspected tuberculous meningitis.

We used the nested polymerase chain reaction (PCR) assays developed previously to detect and identify Mycobacterium tuberculosis (M. tuberculosis) in the cerebrospinal fluid (CSF) samples from patients with suspected tuberculous meningitis and non-tuberculous patients. Our nested PCR assays target the multi-copy IS6110 insertion element and the single-copy mtp40 genomic DNA of M. tuberculosis. These assays, when used in combination, allowed us to detect a very low number of M. tuberculosis in the CSF samples, which otherwise would be undetectable by the culture method, and to distinguish M. tuberculosis from M. bovis. We applied these nested PCR assays to analyze eleven CSF samples. Among these, five of them were from patients with suspected tuberculous meningitis but all except one were culture negative. Our results of PCR assays show that three of these five are M. tuberculosis positive, one of which is M. bovis positive, and only one is M. tuberculosis negative. The other six CSF samples were from the clinically diagnosed non-tuberculous patients. Surprisingly, two of these so called non-tuberculous patients, a subarachnoid hemorrhage (SAH) and the syndrome of inappropriate antidiuretic hormone secretion (SIADH), were shown M. tuberculosis positive. This finding supports a long-standing argument that tuberculous meningitis is one of the causes of these neurological diseases. These nested PCR assays thus provide the neurologists with an important adjunct, in addition to the patient's clinical presentation and laboratory data, for the diagnosis of tuberculous meningitis.

Adolescent↗
PubMed1999

Romano-Ward long QT syndrome: identification of a HERG mutation in a Taiwanese kindred.

Romano-Ward syndrome is an autosomal dominant long-QT syndrome (LQTS) that predisposes affected individuals to sudden death from tachyarrhythmias. We investigated the molecular basis of LQTS in a Taiwanese kindred. Clinical and genetic analyses revealed that a mutation was linked to the human ether-a-go-go-related gene (HERG). The coding sequences and exon-intron borders of HERG were amplified by means of polymerase chain reaction and subjected to single-strand conformation polymorphism (SSCP) analysis. An exon with an aberrant SSCP pattern was cloned and sequenced to study the molecular lesion. A C-->T transition in codon 614, leading to substitution of a valine for an alanine residue in the pore region of the HERG protein, was identified. Analysis with Bsp12861 endonuclease digestion showed the mutation to be present in all affected family members. Given that an unaffected paternal uncle had inherited the same allele from the grandfather as the proband's father, a de novo mutation had apparently occurred in the father and was transmitted to his offspring. In addition to offering presymptomatic genetic diagnosis, identification of the disease-causing mutation may suggest new therapeutic approaches for treatment and prevention of this cardiovascular disease.

Adult↗
PubMed1998

An MTP inhibitor that normalizes atherogenic lipoprotein levels in WHHL rabbits.

Patients with abetalipoproteinemia, a disease caused by defects in the microsomal triglyceride transfer protein (MTP), do not produce apolipoprotein B-containing lipoproteins. It was hypothesized that small molecule inhibitors of MTP would prevent the assembly and secretion of these atherogenic lipoproteins. To test this hypothesis, two compounds identified in a high-throughput screen for MTP inhibitors were used to direct the synthesis of a highly potent MTP inhibitor. This molecule (compound 9) inhibited the production of lipoprotein particles in rodent models and normalized plasma lipoprotein levels in Watanabe-heritable hyperlipidemic (WHHL) rabbits, which are a model for human homozygous familial hypercholesterolemia. These results suggest that compound 9, or derivatives thereof, has potential applications for the therapeutic lowering of atherogenic lipoprotein levels in humans.

Alanine Transaminase↗
PubMed1998

Backbone and side chain dynamics of uncomplexed human adipocyte and muscle fatty acid-binding proteins.

Adipocyte lipid-binding protein (A-LBP) and muscle fatty acid-binding protein (M-FABP) are members of a family of small ( approximately 15 kDa) cytosolic proteins that are involved in the metabolism of fatty acids and other lipid-soluble molecules. Although highly homologous (65%) and structurally very similar, A-LBP and M-FABP display distinct ligand binding characteristics. Since ligand binding may be influenced by intrinsic protein dynamical properties, we have characterized the backbone and side chain dynamics of uncomplexed (apo) human A-LBP and M-FABP. Backbone dynamics were characterized by measurements of 15N T1 and T2 values and ¿1H¿-15N NOEs. These data were analyzed using model-free spectral density functions and reduced spectral density mapping. The dynamics of methyl-containing side chains were charaterized by measurements of 2H T1 and T1rho relaxation times of 13C1H22H groups. The 2H relaxation data were analyzed using the model-free approach. For A-LBP, 15N relaxation data were obtained for 111 residues and 2H relaxation data were obtained for 42 methyl groups. For M-FABP, 15N relaxation data were obtained for 111 residues and 2H relaxation data were obtained for 53 methyl groups. The intrinsic flexibilities of these two proteins are compared, with particular emphasis placed on binding pocket residues. There are a number of distinct dynamical differences among corresponding residues between the two proteins. In particular, many residues display greater backbone picosecond to nanosecond and/or microsecond to millisecond time scale mobility in A-LBP relative to M-FABP, including F57, K58, and most residues in alpha-helix 2 (residues 28-35). Variations in the dynamics of this region may play a role in ligand selectivity. The side chains lining the fatty acid binding pocket display a wide range of motional restriction in both proteins. Side chains showing distinct dynamical differences between the two proteins include those of residues 20, 29, and 51. This information provides a necessary benchmark for determining dynamical changes induced by ligand binding and may ultimately lead to an enhanced understanding of ligand affinity and selectivity among fatty acid-binding proteins.

Adipocytes↗
PubMed1998

Evidence that microsomal triglyceride transfer protein is limiting in the production of apolipoprotein B-containing lipoproteins in hepatic cells.

The microsomal triglyceride transfer protein (MTP) is a heterodimeric lipid transfer protein that is required for the assembly and secretion of apolipoprotein B (apoB)-containing lipoproteins. A key unresolved question is whether the MTP-mediated step is rate limiting. To address this, a unique experimental strategy was used that allowed the in situ modulation and measurement of MTP triglyceride transfer activity. In order to accomplish this, an irreversible photoaffinity inhibitor, BMS-192951, was designed and synthesized. When incubated with purified MTP and irradiated with UV light at 360 nm, BMS-192951 inhibits triglyceride transfer by covalently binding to the protein. HepG2 cells were treated with either increasing concentrations of BMS-192951 (0-15 microM) with 5 min of ultraviolet irradiation, or 3.0 microM BMS-192951 with various lengths (0-15 min) of ultraviolet irradiation. Microsomal extracts were prepared exhaustively dialyzed to remove unbound inhibitor, and assayed for MTP-mediated triglyceride transfer activity. BMS-192951 was shown to reduce MTP activity in both a dose- and UV exposure time-dependent fashion. Measurement of apoB concentration in the media showed that apoB secretion was reduced in proportion to the in situ inhibition of MTP activity, while no change was observed in apoA-I secretion. Experiments performed in McArdle RH-7777 rat hepatoma cells and primary rat hepatocytes gave nearly identical results; the decrease in apoB secretion was proportional to the decrease in MTP activity. These results indicate that MTP-mediated lipid transfer is limiting in the assembly and secretion of apoB-containing lipoproteins in hepatic cells under the conditions tested.

Affinity Labels↗
PubMed1996

An inhibitor of the microsomal triglyceride transfer protein inhibits apoB secretion from HepG2 cells.

The microsomal triglyceride (TG) transfer protein (MTP) is a heterodimeric lipid transfer protein that catalyzes the transport of triglyceride, cholesteryl ester, and phosphatidylcholine between membranes. Previous studies showing that the proximal cause of abetalipoproteinemia is an absence of MTP indicate that MTP function is required for the assembly of the apolipoprotein B (apoB) containing plasma lipoproteins, i.e., very low density lipoproteins and chylomicrons. However, the precise role of MTP in lipoprotein assembly is not known. In this study, the role of MTP in lipoprotein assembly is investigated using an inhibitor of MTP-mediated lipid transport, 2-[1-(3, 3-diphenylpropyl)-4-piperidinyl]-2,3-dihydro-1H-isoindol-1-o ne (BMS-200150). The similarity of the IC50 for inhibition of bovine MTP-mediated TG transfer (0.6 microM) to the Kd for binding of BMS-200150 to bovine MTP (1.3 microM) strongly supports that the inhibition of TG transfer is the result of a direct effect of the compound on MTP. BMS-200150 also inhibits the transfer of phosphatidylcholine, however to a lesser extent (30% at a concentration that almost completely inhibits TG and cholesteryl ester transfer). When BMS-200150 is added to cultured HepG2 cells, a human liver-derived cell line that secretes apoB containing lipoproteins, it inhibits apoB secretion in a concentration dependent manner. These results support the hypothesis that transport of lipid, and in particular, the transport of neutral lipid by MTP, plays a critical role in the assembly of apoB containing lipoproteins.

Animals↗
PubMed1996

Crystallization of microsomal triglyceride transfer protein from bovine liver.

The microsomal triglyceride transfer protein (MTP) is a heterodimeric lipid transfer protein required for the assembly of plasma very low density lipoproteins in the liver and chylomicrons in the intestine. Bovine MTP was purified by a modification of a previously published procedure and crystals of MTP were grown reproducibly with polyethylene glycol as a precipitant at pH 7.0. MTP crystals, which diffract to Bragg spacings of better than 3.2 A, have the symmetry of space group P2(1)2(1)2(1) with refined lattice constants of a = 88.7, b = 100.9 and c = 201.1 A, with one heterodimer per asymmetric unit.

Journal Article↗
PubMed1996

Source localization and bioelectric imaging as it relates to continuous waveform analysis.

One of the goals of the mathematical analysis of scalp-recorded continuous EEG waveforms is to elucidate non-invasively the neural generators of these voltages. One way of accomplishing this is to simulate these generators by equivalent current sources and follow the apparent motion of these theoretical generators during the temporal evolution of the EEG. Another way of accomplishing this is to follow the changes in scalp or simulated cortical surface potential or Laplacian maps during the temporal evolution of the EEG. We first discuss the possible theoretical pitfalls of using linear techniques on an essentially nonlinear problem (the localization of the sources of the EEG), as well as possible computational pitfalls associated with realistic, but complex, conductive medium models simulating the head. Various mathematical source localization methods and bioelectric imaging techniques are then outlined. Later in this paper a collaborative project involving mathematicians, computer scientists, and clinical neuroscientists is described. In this project EEG waveform data will be analyzed, millisecond-to-millisecond, using the various mathematical techniques for localizing equivalent current sources and simulating cortical surface potential and Laplacian topographical maps mentioned above. One of the clinical aims of this research project is to localize epileptic foci without employing invasive recording procedures. Since the EEG datasets that will be analyzed are large (22 Mb, in some cases), data compression and parallel processing strategies will be important parts of this research project. Such strategies, as they may apply to continuous waveform analysis, are discussed in the two appendices at the end of this paper. Many of the presentations at the Symposium and the corresponding papers in this Supplement are related to the ideas and goals of this research project and the implementation of the mathematical/computational techniques for realizing these goals.

Brain↗
PubMed1996

A pilot study of the marginal adaptation and surface morphology of glass-cermet cements.

This study investigated changes in the marginal adaptation and surface morphology of Ketac-Silver and Chelon-Silver glass-current cements over time. Dispersalloy amalgam was used as a control. Contralateral pairs of carious primary molars were restored with the test materials and amalgam. Clinical evaluations were scheduled at 12, 18, and 24 months after placement. Gold-plated replicas of the restorations were observed with scanning electron microscopy. Fractures and cracks in the surface of the Dispersalloy and Chelon-Silver increased the surface roughness; however, the damage was superficial and self-limiting in the Dispersalloy restorations, while in Chelon-Silver the fractures caused the material to break down in layers. A substantial quantity of pores, usually smaller than 50 microns in diameter, were observed throughout the surface of the Chelon-Silver restorations. The pores in the surface of Ketac-Silver were fewer and smaller. The incidence of cavomarginal breakdown increased with time. Chelon-Silver restorations had a higher rate of cavomarginal breakdown than did Ketac-Silver and Dispersalloy restorations up to 18 months. However, there was no statistically significant difference in the marginal adaptation of the three groups at 24 months.

Cermet Cements↗
PubMed1995

Microsomal triglyceride transfer protein. Specificity of lipid binding and transport.

Microsomal triglyceride transfer protein (MTP) is a lipid transfer protein that is required for the assembly and secretion of very low density lipoproteins by the liver and chylomicrons by the intestine. To further elucidate the nature of the lipid molecule binding and transport site on MTP, we have studied the relative rates at which MTP transports different lipid species. Assay conditions were chosen in which there were minimal changes in the physical properties of the substrate membranes so that transfer rates would reflect MTP-lipid interactions at a membrane surface. Lipid transport rates decreased in order of triglyceride > cholesteryl ester > diglyceride > cholesterol > phosphatidylcholine. Changes in the hydrophobic nature of a lipid molecule by the addition of a fatty acid, modulated the ability of MTP to transport it. Addition of one acyl chain from diglyceride to triglyceride, lysophosphatidylcholine to phosphatidylcholine, or cholesterol to cholesteryl ester increased the rate of MTP-mediated transport 10-fold. In contrast, the lipid transport rate was insensitive to the changes in the structure or charge of the polar head group on phospholipid substrates. Zwitterionic, net negative, or net positive charged phospholipid molecules were all transported at a comparable rate. The ability of MTP to transport lipids is strongly correlated to the binding of these lipids to MTP. Thus, MTP has a specific preference for binding and transporting nonpolar lipid compared with phospholipids, and within a class of lipid molecules, a decrease in polarity increases its tendency to be transported.

Animals↗
PubMed1994

Alternative adrenal chromaffin cell fates induced by basic fibroblast growth factor or cyclic AMP in vitro depend on a collaboration with the growth substrate.

Basic fibroblast growth factor stimulates cultured adrenal chromaffin cells to divide and to transform into sympathetic neurons, but the efficacies reported for these actions of basic fibroblast growth factor have varied widely. We have examined the effect of various growth substrates on basic fibroblast growth factor responses and here we show that the ability of basic fibroblast growth factor to transform neonatal rat chromaffin cells into sympathetic neurons depends on laminin as the culture substrate. On collagen, less than 5% of the cells were transformed into neurons by basic fibroblast growth factor, even when the culture was supplemented with heparin or heparan sulphate, but 65% of cells entered the S phase in the presence of basic fibroblast growth factor compared to 15% in its absence, showing that the basic fibroblast growth factor receptor is still active. On laminin, by contrast, over 60% of the cells transformed into neurons in response to the same concentrations of basic fibroblast growth factor, suggesting that an overlapping pool of cells change their phenotype depending on growth substrate. The cyclic AMP analogue 8-(4-chlorophenylthio)cyclic AMP apparently mimicked the actions of basic fibroblast growth factor, promoting neuronal differentiation on laminin, but mitogenic stimulation on collagen. These data support the notion that collagen and laminin promote different instructions in chromaffin cells that can collaborate with the signals induced by basic fibroblast growth factor and 8-(4-chlorophenylthio)cyclic AMP to determine chromaffin or neuronal cell fates.

Adrenal Glands↗
PubMed1993

Effects of histamine on intracellular Na+ activity and twitch tension in guinea pig papillary muscles.

Effects of histamine on the twitch tension, intracellular Na+ activity (aiNa), and action potential were simultaneously and continuously measured in the isolated cardiac papillary muscle of guinea pigs. In the driven fiber at 60 beats/ min, histamine induced a concentration-dependent positive inotropic effect, and apparently increased the amplitude of the action potential and shortened the duration at 3 microM, and significantly decreased aiNa by 6.7% from the control value of 4.5 +/- 0.1 mM (n = 6, p < 0.05) at 0.3 microM. Cimetidine at 10 microM did not produce any significant effect but partially antagonized the action of histamine. The respective concentrations of 1 and 3 microM histamine significantly decreased aiNa by 5.4 and 14.3% from the control 5.6 +/- 0.1 mM (n = 6, p < 0.05) in the presence of 10 microM cimetidine. Chlorpheniramine at 10 microM significantly reduced the twitch tension and aiNa, and prolonged the action potential duration. Histamine at 1 microM increased the twitch tension and decreased aiNa from 4.5 +/- 0.1 to 4.1 +/- 0.1 mM (n = 5, p < 0.05) in the presence of 10 microM chlorpheniramine. The combined application of 10 microM cimetidine and 10 microM chlorpheniramine reduced the twitch tension and abolished most effects of histamine. In high potassium solution (16.2 mM), 1 microM histamine significantly decreased aiNa by 12.9% from the control of 3.1 +/- 0.2 mM (n = 5, p < 0.05). The treatment of 0.5 or 1 microM ouabain increased the tension development and aiNa, and abolished the decrease in aiNa produced by the effect of histamine. The results suggest that histamine can directly stimulate Na-K pump in sarcolemma through means of the activation of both H1- and H2-receptors.

Action Potentials↗
PubMed1990

Nonlinear methods in electrocardiogram signal processing.

Electrocardiographic (ECG) signals are frequently corrupted by impulsive noise due to muscle activities, and background normalization is often needed to correct for patient motion and respiration. Nonlinear signal processing methods are effective alternatives to conventional linear filtering methods when dealing with impulsive noise or noise types that are difficult to characterize. The class of nonlinear filtering methods studied in this article operate by moving a window of finite width along the input data sequence. At each position, the filter output is obtained from the input samples inside the window. Nonlinear operators differ from linear filters in that the output is not a simple linear combination of the input samples. Three classes of nonlinear operators--median filters, morphologic operators, and the alpha-trimmed mean filter--are briefly introduced and algorithms using them for ECG signal processing are presented. Empirical results indicate that the nonlinear operators are good candidates for impulsive noise suppression and background normalization in ECG signal processing.

Algorithms↗
PubMed1989

The 3-hydroxyacyl-CoA epimerase activity of rat liver peroxisomes is due to the combined actions of two enoyl-CoA hydratases: a revision of the epimerase-dependent pathway of unsaturated fatty acid oxidation.

Chromatography of a rat liver extract on DEAE-cellulose resulted in the near total loss of 3-hydroxyacyl-CoA epimerase activity. The activity was regained either when fractions were recombined or when purified crotonase was added to the early column fractions. A new enoyl-CoA hydratase present in these early fractions catalyzes the conversion of D-3-hydroxyacyl-CoA to 2-trans-enoyl-CoA which can be hydrated by crotonase or the peroxisomal bifunctional enzyme to L-3-hydroxyacyl-CoA. Thus, the 3-hydroxyacyl-CoA epimerase activity is due to the combined actions of two enoyl-CoA hydratases with opposite stereospecificities.

Animals↗
PubMed1989

Impulsive noise suppression and background normalization of electrocardiogram signals using morphological operators.

A new approach to impulsive noise suppression and background normalization of digitized electrocardiogram signals is presented using mathematical morphological operators that incorporate the shape information of a signal. A brief introduction to these nonlinear signal processing operators, as well as a detailed description of the new algorithm, is presented. Empirical results show that the new algorithm has good performance in impulsive noise suppression and background normalization.

Algorithms↗
PubMed1986

Mailable TLD system for photon and electron therapy beams.

The mailable TLD system developed by the Radiological Physics Center for monitoring calibration of photon beam energies from cobalt 60 to 25 MV and electron beam energies from 6 to 20 MeV has been in use since 1977 for photons and since 1982 for electron beams. Design considerations, proper use of the system and calibration techniques are detailed. The accuracy of the system is comparable to that of ion chamber measurements made in a water phantom, although it shows less precision.

Electrons↗
PubMed1985

3-Hydroxyacyl-CoA epimerase is a peroxisomal enzyme and therefore not involved in mitochondrial fatty acid oxidation.

The subcellular location of 3-hydroxyacyl-CoA epimerase (EC 5.1.2.3) was studied by differential centrifugation and Percoll density gradient centrifugation of a rat liver homogenate. The enzyme was found to be associated with peroxisomes but not with mitochondria. This observation proves that 3-hydroxy-acyl-CoA epimerase does not function in mitochondrial beta-oxidation of polyunsaturated fatty acids which are degraded by a modified pathway.

Animals↗