Biomedical subjects

D H Chen

Publications and source records attributed to D H Chen.

At least 19 recordsLinked to original sources

PubMed2001

Endostatin binds tropomyosin. A potential modulator of the antitumor activity of endostatin.

The mechanism of action of Endostatin, an endogenous inhibitor of angiogenesis and tumor growth, remains unknown. We utilized phage-display technology to identify polypeptides that mimic the binding domains of proteins with which Endostatin interacts. A conformed peptide (E37) was identified that shares an epitope with human tropomyosin implicating tropomyosin as an Endostatin-binding protein. We show that recombinant human Endostatin binds tropomyosin in vitro and to tropomyosin-associated microfilaments in a variety of endothelial cell types. The most compelling evidence that tropomyosin modulates the activity of Endostatin was demonstrated when E37 blocked greater than 84% of the tumor-growth inhibitory activity of Endostatin in the B16-BL6 metastatic melanoma model. We conclude that the E37 peptide mimics the Endostatin-binding epitope of tropomyosin and blocks the antitumor activity of Endostatin by competing for Endostatin binding. We postulate that the Endostatin interaction with tropomyosin results in disruption of microfilament integrity leading to inhibition of cell motility, induction of apoptosis, and ultimately inhibition of tumor growth.

Actin Cytoskeleton↗

PubMed2001

Both retinoic acid and 1,25(OH)2 vitamin D3 inhibit thyroid hormone-induced terminal differentiaton of growth plate chondrocytes.

Thyroid hormone has been known for over 50 years to be a potent regulator of skeletal maturation at the growth plate. The receptor for thyroid hormone has been discovered to be a member of the nuclear hormone receptor superfamily. Retinoic acid and 1,25(OH)2 vitamin D3, whose receptors also belong to this nuclear hormone receptor family, have been implicated in the control of chondrocyte proliferation and differentiation at the growth plate. Recent studies demonstrate that the receptors for thyroid hormone, retinoic acid, and vitamin D bind to a similar DNA response element in the promoter region of target genes and may form heterodimers to regulate gene transcription in target cells. These observations led us to hypothesize that the retinoic acid and/or vitamin D signaling pathways may interact with thyroid hormone signaling at the molecular level to modulate growth plate chondrocyte differentiation. Using a chemically defined, serum-free model of growth plate chondrocyte maturation, both all-trans retinoic acid and 1,25(OH)2 vitamin D3 markedly inhibited thyroid hormone-induced terminal differentiation in a dose-dependent manner. In the absence of thyroid hormone, retinoic acid stimulated alkaline phosphatase activity modestly at the highest dose used, however neither retinoic acid nor 1,25(OH)2 vitamin D3 induced expression of type X collagen mRNA. We conclude that retinoic acid and vitamin D are likely to be antagonists of thyroid hormone signaling in the growth plate.

Animals↗

PubMed2001

Chemical constituents of Typhonium giganteum Engl.

A new cerebroside, named typhonoside (1), was isolated from the root tuber of Typhonium giganteum Engl. along with three known compounds dipalmitin (2), alpha-monopalmitin (3) and 2,6-diamino-9-beta-D-ribofuranosylpurine (4). The structure of 1 was determined to be 1-O-beta-D-glucopyranosyl-(2S,3S,4R,8Z)-2-[(2'-hydroxyl-docosanoyl)amino]-8-otadecene-1,3,4-triol on the basis of spectral data.

Cerebrosides↗

PubMed2001

The pattern of tegument-capsid interaction in the herpes simplex virus type 1 virion is not influenced by the small hexon-associated protein VP26.

Examination of the three-dimensional structure of intact herpes simplex virus type 1 (HSV-1) virions had revealed that the icosahedrally symmetrical interaction between the tegument and capsid involves the pentons but not the hexons (Z. H. Zhou, D. H. Chen, J. Jakana, F. J. Rixon, and W. Chiu, J. Virol. 73:3210-3218, 1999). To account for this, we postulated that the presence of the small capsid protein, VP26, on top of the hexons was masking potential binding sites and preventing tegument attachment. We have now tested this hypothesis by determining the structure of virions lacking VP26. Apart from the obvious absence of VP26 from the capsids, the structures of the VP26 minus and wild-type virions were essentially identical. Notably, they showed the same tegument attachment patterns, thereby demonstrating that VP26 is not responsible for the divergent tegument binding properties of pentons and hexons.

Base Sequence↗

PubMed2001

[Thermal decomposition kinetics of ribavirin and its stability].

AIM: To study Thermal stabilities and thermal decomposition process of ribavirin and establish thermal decomposition kinetics equation. METHODS: Thermal weight loss curve was obtained by thermogravimetry balance. Thermal decomposition function mode was also determined by Achar differential method and Coats-Redfern integral method. RESULTS: Thermal decomposition kinetics parameters and kinetics compensation parameters were calculated from thermogravimetry-differential thermogravimetry data. Activation energy deduced by extrapolation under heating rate of 0 degree C.min-1 was 188.04 kJ.mol-1. Thermal decomposition kinetics function expression was d alpha/dt = Ae [formula: see text] (1 - alpha)2 and the mathematic expression of the kinetic compensation effect is found to be InA = 0.2264 Ea - 5.4458. CONCLUSION: Ribavirin has high thermal stability because of its high thermal decomposition activation energy. The thermal decomposition activation of capsule is little lower than that of material, which indicates no evident difference in thermal stability for capsules and material.

Antiviral Agents↗

PubMed2000

Expression of cyclin-dependent kinase inhibitors in epiphyseal chondrocytes induced to terminally differentiate with thyroid hormone.

A growing body of evidence suggests that systemic hormones and peptide growth factors may exert their effects on cell growth and differentiation in part through regulation of the cell division cycle. We hypothesized that thyroid hormone regulates terminal differentiation of growth plate chondrocytes in part through controlling cell cycle progression at the G1/S restriction point. Our results support this hypothesis by demonstrating that treatment of epiphyseal chondrocytes with thyroid hormone under chemically defined conditions results in the arrest of DNA synthesis and the onset of terminal differentiation, indicating that thyroid hormone is one factor capable of regulating the transition between cell growth and differentiation in these cells. This terminal differentiation process is associated with induction of the cyclin/cyclin-dependent kinase inhibitors p21(cip-1 waf-1) and p27kip1, suggesting that thyroid hormone may regulate terminal differentiation in part by arresting cell cycle progression through induction of cyclin-dependent kinase inhibitors.

Animals↗

PubMed1999

Three-dimensional visualization of tegument/capsid interactions in the intact human cytomegalovirus.

The three-dimensional structure of the intact human cytomegalovirus (HCMV) was determined to 18-A resolution by electron cryomicroscopy and computer reconstruction. Its capsid shell is composed of pentons, hexons, and triplexes arranged on a T = 16 icosahedral lattice and is identical to that of the B-capsid isolated from host cell nuclei. An icosahedrally ordered tegument layer formed by 960 copies of filamentous density is also visualized, which interacts with the pentons, hexons, and triplexes of the underlying capsid. The observed structural similarities and differences of HCMV with those of herpes simplex virus offer insights into the significance of the different tegument components for their infection processes while maintaining similar capsids.

Capsid↗

PubMed1999

Visualization of protein-RNA interactions in cytoplasmic polyhedrosis virus.

Unlike the multiple-shelled organization of other Reoviridae members, cytoplasmic polyhedrosis virus (CPV) has a single-shelled capsid. The three-dimensional structures of full and empty CPV by electron cryomicroscopy show identical outer shells but differ inside. The outer surface reveals a T=1 icosahedral shell decorated with spikes at its icosahedral vertices. The internal space of the empty CPV is unoccupied except for 12 mushroom-shaped densities attributed to the transcriptional enzyme complexes. The ordered double-stranded RNA inside the full capsid forms spherical shells spaced 25 A apart. The RNA-protein interactions suggest a mechanism for RNA transcription and release.

Animals↗

PubMed1999

Visualization of tegument-capsid interactions and DNA in intact herpes simplex virus type 1 virions.

Herpes simplex virus type 1 virions were examined by electron cryomicroscopy, allowing the three-dimensional structure of the infectious particle to be visualized for the first time. The capsid shell is identical to that of B-capsids purified from the host cell nucleus, with the exception of the penton channel, which is closed. The double-stranded DNA genome is organized as regularly spaced ( approximately 26 A) concentric layers inside the capsid. This pattern suggests a spool model for DNA packaging, similar to that for some bacteriophages. The bulk of the tegument is not icosahedrally ordered. However, a small portion appears as filamentous structures around the pentons, interacting extensively with the capsid. Their locations and interactions suggest possible roles for the tegument proteins in regulating DNA transport through the penton channel and binding to cellular transport proteins during viral infection.

Capsid↗

PubMed1999

A novel deletion of the dystrophin S-promoter region cosegregating with mental retardation.

A contiguous deletion of the S-promoter/first S exon and its downstream exon 56 of the dystrophin gene was identified in a Japanese dystrophinopathy family in which two brothers and their half brother were affected. Characteristically, severe mental retardation cosegregated with the deletion even though they grew up in different environments. Furthermore, mild cerebral atrophy was observed by CT scan and MRI in the eldest brother.

Adult↗

PubMed1999

Expression of thyroid hormone receptor isoforms in rat growth plate cartilage in vivo.

Although thyroid hormone has been known for many years to be a potent regulator of skeletal maturation in vivo, it has not definitively been determined whether this effect is a result of a direct or indirect action of the hormone. Previous in vivo studies have suggested that thyroid hormone may stimulate longitudinal bone growth by increasing the secretion of growth hormone; however, growth hormone alone is unable to stimulate cartilage maturation. There are also indications that thyroid hormone is able to act directly on growth plate chondrocytes through growth hormone-independent mechanisms. In this study, we demonstrate that rat growth plate chondrocytes in vivo express genes encoding three of the four isoforms of the thyroid hormone receptors described to date, but the corresponding protein can only be detected for the TRalpha1 and TRbeta1 isoforms of the receptor. As has been noted in other tissues, there is generally poor correlation between the mRNA levels for each isoform and the relative amount of corresponding protein as measured by immunoblotting, suggesting the possibility that receptor expression may be regulated by post-transcriptional mechanisms.

Animals↗

PubMed1999

Urinary tract infection in children.

Two hundred and eighty-five children under 15 years old (169 boys and 116 girls) who had urinary tract infection (UTI) were admitted to our hospital during the period July 1995 to June 1998. Clinical presentation, laboratory data and image studies were recognized and analyzed. Most patients were younger than 2 years of age. Fever is the most common sign, especially in young children. With the urinary dipstick test a low positive rate of nitrite reaction (34.7%) was found. E. coli is the most common pathogen and exclusively resistant to ampicillin (90.2%). The E. coli is also relatively resistant to trimethoprim/sulfamethoxazole (57.1%). Voiding cystourethrogram was performed in 187 cases, in which 24.6% of those patients showed vesicoureteric reflux. Ninety-eight children received 99mTc-dimercaptosuccinic acid renal scans and 62 cases (63.3%) had abnormal findings compatible with pyelonephritis. The positive predictive values by renal ultrasonography for vesicoureteric reflux and pyelonephritis are 30.7% and 78.9%, respectively. In pyelonephritis patients, only 37% also had a vesicoureteric reflux. The fever duration and clinical inflammatory parameters were evaluated in all patients. Longer febrile periods are the risk of vesicoureteric reflux and pyelonephritis, and a high C-reactive protein concentration indicates the risk of pyelonephritis. In conclusion, fever was the most common sign in young UTI children who had a low positive nitrite reaction rate in the dipstick test. E. coli was the most common pathogen with a high ampicillin resistance in Taiwan. Only 37% pyelonephritis patients had refluxing nephropathy. It is indicated that pyelonephritis is not always attributed to reflux of infected urine. A longer febrile period and a high C-reactive protein level are good indicators for prediction of the risk of pyelonephritis in UTI patients.

Adolescent↗

PubMed1998

Characterization of p96h2bk: immunoreaction with an anti-Erk(extracellular-signal-regulated kinase) peptide antibody and activity in Xenopus oocytes and eggs.

We have shown previously that oncogenic Ras induces cell cycle arrest in activated Xenopus egg extracts [Pan, Chen and Lin (1994) J. Biol. Chem. 269, 5968-5975]. The cell cycle arrest correlates with the stimulation of a protein kinase activity that phosphorylates histone H2b in vitro (designated p96(h2bk)) [Chen and Pan (1994) J. Biol. Chem. 269, 28034-28043]. We report here that p96(h2bk) is likely to be p96(ram), a protein of approx. 96 kDa that immunoreacts with a monoclonal antibody (Mk-1) raised against a synthetic peptide derived from a sequence highly conserved in Erk1/Erk2 (where Erk is extracellular-signal-regulated kinase). This is supported by two lines of evidence. First, activation/inactivation of p96(h2bk) correlates with upward/downward bandshifts of p96(ram) in polyacrylamide gels. Secondly, both p96(h2bk) and p96(ram) can be immunoprecipitated by antibody Mk-1. We also studied the activity of p96(h2bk)/p96(ram) in Xenopus oocytes and eggs. p96(h2bk)/p96(ram) was inactive in stage 6 oocytes, was active in unfertilized eggs, and became inactive again in eggs after fertilization. Since stage 6 oocytes are at G2-phase of the cell cycle, unfertilized eggs arrest at M-phase and eggs exit M-phase arrest after fertilization, the results thus indicate that p96(h2bk)/p96(ram) activity is cell cycle dependent. Moreover, microinjection of oncogenic Ras into fertilized eggs at the one-cell stage arrests the embryos at the two-cell stage, and this induced arrest is correlated with an inappropriate activation of p96(h2bk)/p96(ram). The data are consistent with the concept that inappropriate activation of p96(h2bk)/p96(ram) plays a role in the cell cycle arrest induced by oncogenic Ras.

Animals↗

PubMed1997

The 96 kDa protein kinase activated by oncogenic Ras in Xenopus egg extracts is also activated by constitutively active Mek: activation requires serine/threonine phosphorylation.

In the Xenopus egg and oocyte system, oncogenic Ras protein can induce cell cycle arrest. The effect of oncogenic Ras on the cell cycle seems to be mediated by the Raf-Mek-Erk pathway of Ras signal transduction since constitutively active Raf, Mek, or Erk can mimic the effect of oncogenic Ras protein and since specific inhibition of these kinases can block the effect of oncogenic Ras. Using activated Xenopus egg extracts, we previously reported that the cell cycle arrest induced by oncogenic Ras correlates with the activation of a 96 kDa protein that phosphorylates histone h2b in vitro. This result raised the possibility that the 96 kDa kinase (designated as p96h2bk) is a potential target of the Raf-Mek-Erk pathway that links the pathway to the control of the cell cycle. We report here that constitutively active Mek1 could activate p96h2bk in the absence of oncogenic Ras. Moreover, inhibition of endogenous Mek by a specific inhibitor, PD 098059, suppressed the activation of p96h2bk by oncogenic Ras. These results are consistent with the concept that p96h2bk is a component or target of the Raf-Mek-Erk pathway. Furthermore, we have shown that activation of p96h2bk requires serine/threonine phosphorylation of p96h2bk.

Animals↗

PubMed1997

Ultrastructural changes in rat locus coeruleus induced by chronic opioids.

The locus coeruleus (LC) is a major noradrenergic nucleus in the brain. The activity of the LC neurons is chronically regulated by opioids. So far, very little is known about the morphological changes induced by chronic treatment with opioids. In the present study, the effects of chronic treatment with morphine and dihydroetorphine, a new narcotic analgesic with lower physical dependence potential than morphine, were investigated on the ultrastructure of the rat LC. Rats received saline or increasing doses of morphine or dihydroetorphine for 5 days by twice daily subcutaneous injections. Withdrawal was precipitated in half of the opioid-treated rats by a single intraperitoneal injection of naloxone 4 h after the last injections of opioids. The ultrastructure of the LC was examined by electron microscopy. Results showed that chronic morphine treatment induced a marked injury to the LC neurons. The primary changes in the cell body were the indentation of nuclei, the fragmentation and degranulation of rough endoplasmic reticulum, as well as the disaggregation of polyribosomes. Myelinoid bodies were seen in the processes. An accumulation of presynaptic vesicles was observed in some of the terminals which formed synaptic junctions with the LC neurons as compared to the normal controls. Naloxone-precipitated withdrawal from morphine did not stop the morphine-induced injury on the LC neurons except that less accumulation of presynaptic vesicles occurred. Chronic dihydroetorphine treatment only induced a slight change in the ultrastructure of the LC neurons. These results indicate that the LC neurons are more vulnerable to chronic treatment with morphine than to that with dihydroetorphine.

Analgesics↗

PubMed1996

Identification of a potential physiological substrate for oncogenic Ras-activated protein kinases in activated Xenopus egg extracts: correlation with oncogenic Ras-induced cell cycle arrest.

Activated Xenopus egg extracts are capable of undergoing cell-free cell cycling. Using these activated extracts, we previously showed that purified, bacterially expressed oncogenic human RasH protein arrests cell cycle progression. Because oncogenic Ras activates many serine/threonine protein kinases in Xenopus oocytes and egg extracts, it is possible that induction of cell cycle arrest involves the action of oncogenic Ras-activated kinases. Thus, the identification of the physiological substrates for oncogenic Ras-activated kinases is important for elucidating the molecular mechanism underlying oncogenic Ras-induced cell cycle arrest. We used 32P-orthophosphate as a label to identify the potential substrates. Our results demonstrated that the 32P-labeling of both a 32 and a 33 kDa protein were greatly enhanced by oncogenic Ras during the incubation of activated Xenopus egg extracts. The enhanced labeling correlated with the induced cell cycle arrest and was contributed by serine phosphorylation. Moreover, the 33 kDa protein was detected only in the presence of oncogenic Ras and was a serine-hyperphosphorylated form of the 32 kDa protein. Furthermore, new protein synthesis was not required for the enhanced labeling, consistent with the concept that the enhanced serine phosphorylation of the 32 kDa protein is by oncogenic Ras-activated protein kinases. In addition to serine phosphorylation, our results also suggested that an as yet unidentified modification of the 32 kDa protein might also be induced by oncogenic Ras. Our results suggest that the 32 kDa protein is a potential physiological substrate for oncogenic Ras-activated protein kinases.

14-3-3 Proteins↗

PubMed1995

The anticoagulation factor protein S and its relative, Gas6, are ligands for the Tyro 3/Axl family of receptor tyrosine kinases.

We report the identification of ligands for Tyro 3 (alternatively called Sky, rse, brt, or tif) and Axl (alternatively, Ark or UFO), members of a previously orphan family of receptor-like tyrosine kinases. These ligands correspond to protein S, a protease regulator that is a potent anticoagulant, and Gas6, a protein related to protein S but lacking any known function. Our results are reminiscent of recent findings that the procoagulant thrombin, a protease that drives clot formation by cleaving fibrinogen to form fibrin, also binds and activates intracellular signaling via a G protein-coupled cell surface receptor. Proteases and protease regulators that also activate specific cell surface receptors may serve to integrate coagulation with associated cellular responses required for tissue repair and growth, as well as to coordinate protease cascades and associated cellular responses in other systems, such as those involved in growth and remodeling of the nervous system.

3T3 Cells↗

PubMed1994

Transport and hydrolysis of urea in a reactor-separator combining an anion-exchange membrane and immobilized urease.

A membrane reactor-separator, in which an anion-exchange membrane and a urease-immobilized poly(vinyl alcohol) (PVA) membrane were clamped together to separate the feed solution and the stripping solution of a dialysis cell, was constructed. The urea in the feed solution passed through the anion-exchange membrane, water film, and then was hydrolyzed to ammonium carbamate in the urease-immobilized PVA membrane. The experimental results showed that no ammonium ion was found in the feed solution under either phosphate or citrate buffer systems at 0.05-0.2 mol dm-3 and pH 6-9, and various initial concentrations of urea in the feed solution (20-200 mmol dm-3). This indicates that the water film between two membranes allows the carbamate ions to decompose into ammonium and carbonate ions completely before entering the anion-exchange membrane. The device therefore can be used for the removal of urea from feed solution, while preventing the backflow of ammonium ions from the stripping solution or water film into feed solution. It has significant potential in the development of a wearable or portable artificial kidney. The properties of the urease-immobilized PVA membrane were examined. A kinetic model describing the transport-reaction behavior of urea in the membrane reactor-separator was developed, and the optimum values of the reactor parameters were obtained.

Biological Transport↗