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Biomedical subjects

W Chen

Publications and source records attributed to W Chen.

At least 793 records · Page 44Linked to original sources
PubMed1996

Synthesis, chromatographic resolution, and anti-human immunodeficiency virus activity of (+/-)-calanolide A and its enantiomers.

The anti-HIV agent (+/-)-calanolide A (1) has been synthesized in a five-step approach starting with phloroglucinol [-->5-->6-->11-->18-->(+/-)-1], which includes Pechmann reaction, Friedel-Crafts acylation, chromenylation with 4,4-dimethoxy-2-methylbutan-2-ol, cyclization, and Luche reduction. Cyclization of chromene 11 to chromanone 18 was achieved by employing either acetaldehyde diethyl acetal or paraldehyde in the presence of trifluoroacetic acid and pyridine or PPTS. Luche reduction of chromanone 18 at lower temperature preferably yielded (+/-)-1. Reduction of chromone 12, synthesized by Kostanecki-Robinson reaction from chromene 11, failed to afford (+/-)-1. The synthetic (+/-)-1 has been chromatographically resolved into its optically active forms, (+)- and (-)-1. The anti-HIV activities for synthetic (+/-)-1, as well as resultant (+)- and (-)-1, have been determined. Only (+)-1 accounted for anti-HIV activity, which was similar to the data reported for the natural product, and (-)-1 was inactive.

Antiviral Agents↗
PubMed1996

Adhesion mediated by fibronectin's alternatively spliced EDb (EIIIB) and its neighboring type III repeats.

Adhesion functions of cellular fibronectin's (FN) alternatively spliced EDb (EIIIB) domain, as well as its neighboring type III repeats III7 and III8, were investigated with several cultured murine and human cell types. Minigene constructs encoding various permutations of these repeats and expressed in bacteria were used as shown previously in function studies of EDa and its neighboring repeats (P.Xia and L. A. Culp Exp. Cell Res. 213, 253-265, 1994). When substrata of recombinant proteins were incubated with several fibroblastic or neuronal derivative cell lines, cell attachment responses varied widely in a cell-type-specific manner. Balb/c 3T3 cells were shown to adhere to recombinant protein substrata in dose-dependent and EDTA-sensitive manners. Responses also varied with which repeat combinations were being tested, from excellent (Balb/c 3T3, src-3T3), to intermediate (Platt cells), to poor (LZEJ, VA-13, and F11), with EDb plus-containing proteins generally giving better adhesion than EDb minus proteins. On select recombinant proteins, cells showed limited cytoplasmic spreading (3T3 and src-3T3) or neurite extension (Platt and F11), while other cell lines (VA-13 and LZEJ) did not show any morphological changes beyond attachment. Again, EDb plus-containing recombinants were more effective at inducing these morphological changes than the neighboring repeats. These results demonstrate that the EDb domain of cellular FNs and its neighboring type III homology repeats contain important adhesion-promoting sequences, which may be regulated by cells through alternative splicing of FN's primary transcript.

Alternative Splicing↗
PubMed1996

An integrated physical map covering 25 cM of human chromosome 8.

We have developed an integrated physical map for human chromosome 8q23-q24.1 based on STS content analysis of somatic cell hybrids and YAC contigs. Among the 63 STSs used are markers from multiple genetic maps and several previously unmapped polymorphisms, thus integrating genetic markers from different sources into comprehensive, ordered sets based on their positions in the physical map. These maps will facilitate the isolation of genes causing human inherited diseases that map to these chromosomal regions.

Chromosome Mapping↗
PubMed1996

Molecular cloning and characterization of a putative novel human osteoclast-specific 116-kDa vacuolar proton pump subunit.

A cDNA encoding a possible novel human 116-kDa polypeptide subunit of the osteoclastic proton pump (OC-116KDa) has been identified by differential screening of a human osteoclastoma cDNA library. The predicted sequence of OC-116kDa consists of 822 amino acids and is 46.9% and 47.2% identical at the amino acid level to the 116-KDa polypeptide of the vacuolar proton pump of rat and bovine brain respectively. OC-116KDa mRNA was found at high levels in osteoclastomas by Northern analysis but was not detected in tumor stromal cells or in other tissues including kidney, liver, skeletal muscle and brain. OC-116KDa mRNA was localized to multinucleated giant cells within the osteoclastoma tumor by in situ hybridization.

Amino Acid Sequence↗
PubMed1996

Hepatic overexpression of insulin-like growth factor-II in adulthood increases basal and insulin-stimulated glucose disposal in conscious mice.

The physiological role of circulating insulin-like growth factor-II (IGF-II) in adult humans is poorly understood. We recently generated an IGF-II transgenic murine model of persistent IGF-II production (plasma IGF-II approximately 30-fold increased above normal) through over-expression of the transgene driven by the major urinary protein promoter (Rinderknecht, E., and Humbel, R. E. (1978) J. Biol. Chem. 269, 13779-13784). To determine whether in vivo insulin action is improved in these transgenic mice, we performed euglycemic insulin (18 milliunits/kg.min) clamp studies in conscious IGF-II transgenic and in age- and weight-matched control mice. Plasma glucose and insulin concentrations were significantly lower in the IGF-II transgenic compared with both control grouoff Despite decreased plasma glucose concentration, basal hepatic glucose production (HGP) and glucose clearance were increased. During the insulin clamp studies in IGF-II transgenic mice compared with control mice (a) the rates of glucose infusion and glucose uptake were increased by approximately by 65 and approximately 55%, respectively; (b) glycolysis was increased by approximately 12% while glycogen synthesis was approximately 2-fold higher; (c) while the suppression of plasma free fatty acid was similar, the increment in plasma lactate concentration was significantly higher; (d) although HGP was similarly inhibited by insulin, phosphoenolpyruvate gluconeogenesis was enhanced and accounted for a larger portion of HGP (64% versus approximately 40% in control mice). Our data suggest that the persistence of circulating IGF-II in adult mice to levels commonly observed in adult humans (50-70 nM) causes a marked improvement in peripheral (skeletal muscle) insulin action, which is not due to changes in body composition. These results suggest that circulating IGF-II may exert a regulatory role on insulin sensitivity and body composition in humans.

Adult↗
PubMed1996

Ex vivo model of leukocyte migration into herpes simplex virus-infected mouse corneas.

We are investigating neutrophilic infiltration into herpes simplex virus type 1 (HSV-1)-infected mouse corneas. Using a novel ex vivo corneal migration assay, we demonstrated two waves of neutrophil chemotaxis in HSV-1-infected corneas. An early chemotactic gradient developed within 48 h in concert with the appearance of HSV-1 epithelial lesions, peaked by 4 days post-infection (p.i.), and degraded by 6-8 days p.i.. A second chemotactic gradient appeared 10 days after infection, just before the initiation of chronic inflammation. The gradient was maintained in corneas that developed inflammation but rapidly degraded in corneas that failed to develop inflammation. The early chemotactic gradient was established in the absence of T lymphocytes, but the second chemotactic gradient was virtually abrogated by T cell depletion. We conclude that HSV-1 infection induces two temporally separated neutrophil chemotactic gradients in the mouse cornea: an early, transient, T cell-independent gradient; and a later, chronic, T cell-dependent gradient.

Animals↗
PubMed1996

Effect of ischemic preconditioning and PKC activation on acidification during ischemia in rat heart.

Ischemic preconditioning (PC) has been shown to attenuate intracellular acidification during a subsequent period of ischemia, to minimize stunning, and to decrease infarct size, PKC activation has been suggested to be involved in this phenomenon. The present study is designed to test whether PKC activation could mimic and PKC inhibition could block the PC effects on intracellular acidification during ischemia and on stunning during reflow in Langendorff perfused rat hearts. Prior to 20 min of sustained global normothermic ischemia, groups of hearts were treated with the PKC activators 4 beta-phorbol 12-myristate 13-acetate (PMA) or 1,2-dioctanoyl-srt-glycerol (DOG), a group of hearts was treated with the PKC inhibitor chelerythrine (CH), a group was treated with DOG plus CH, a group was preconditioned with four cycles of 5 min of ischemia and 5 min of reflow, and a group was treated with CH during PC. Recovery of left ventricular developed pressure (% of initial, pretreatment, preischemic LVDP), measured after 20 min of reflow, was improved in hearts treated with DOG, but not PMA (80 +/- 3% (DOG), 55 +/- 3% (PMA) v 51 +/- 3% (control), P < 0.05 between DOG and control), although both caused a similar degree of PKC translocation (measured by fractionation followed by an assay of PKC activity using incorporation of 32P into histone). The improved recovery of LVDP in the PC group and in the DOG group was blocked by chelerythrine. Measurement of pH (by 31P NMR) showed that DOG reduced acidification at 15-20 min of ischemia, although the effect was not as great as PC, while PMA did not reduce acidification. The effect of DOG on pHi was attenuated by CH; however, the PC-induced attenuation of the fall in pHi, was not affected by CH. High energy phosphates (measured by 31P NMR) were not significantly different between any of the groups during ischemia or reflow. This study confirms that the protective effect of ischemic preconditioning on stunning in rat heart can be eliminated by inhibition of PKC, but suggests that the effect of PC on the fall in pHi during sustained ischemia is not mediated by PKC.

Animals↗
PubMed1996

A comparison of AMP degradation in the perfused rat heart during 2-deoxy-D-glucose perfusion and anoxia. Part I: The release of adenosine and inosine.

AMP degradation is studied in two models of the Langendorff-perfused rat heart which generate a large release of purines: the 2-deoxy-D-glucose (2DG)-perfused heart and the anoxic heart. In the 2DG model, mitochondrial energy generation is quasi-normal, despite a very low ATP concentration. Furthermore, inorganic phosphate (Pi) concentration is low, an important difference with anoxia where Pi is very high, up to 82 mM. Coronary release of purines is measured by high performance liquid chromatography, and myocardial metabolite content by 31P nuclear magnetic resonance spectroscopy. In the 2DG-perfused hearts with glucose or acetate, the purine release consists nearly exclusively of inosine [up to 130 nmol/(min x gww)] while adenosine is less than 1 nmol/(min x gww). A possible interpretation is that AMP degradation proceeds mainly through deamination to inosine monophosphate by AMP deaminase (the IMP pathway). In contrast, the purine release in anoxia (100% N2) contains comparable quantities of adenosine and inosine [respectively 30 and 20 nmol/(min x gww)], indicating that part of AMP is dephosphorylated directly to adenosine. Comparison with the 2DG model suggests that the release of adenosine in the anoxic heart is a result of inhibition of AMP deaminase by Pi.

Adenosine↗
PubMed1996

AMP degradation in the perfused rat heart during 2-deoxy-D-glucose perfusion and anoxia. Part II: The determination of the degradation pathways using an adenosine deaminase inhibitor.

Using the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA), we determine the contribution of the adenosine pathway to the abundant purine release of two Langendroff-perfused rat heart models which differ particularly in inorganic phosphate (Pi) content: the 2-deoxy-D-glucose (2DG) perfused heart and the anoxic heart. We measure the release of coronary purines by high performance liquid chromatography, and the content of myocardial metabolites by 31P nuclear magnetic resonance spectroscopy. In the 2DG-perfused heart (2 mM for 45 min), the release of inosine [130 nmol/(min.gww)] is much larger than that of adenosine, and EHNA (50 microM) has little effect, showing that the pathway of inosine monophosphate (IMP) accounts for 97% of purine catabolism. In the anoxic heart (100% N2 for 45 min), where inosine and adenosine release are comparable in the absence of EHNA, the inhibitor reduces the release of inosine and catabolites from 50 to 20 nmol/(min.gww) and increases that of adenosine [from 30 to 55 nmol/(min.gww)], showing that the contributions of the IMP and adenosine pathways are 23 and 77%. The difference between the two models has been ascribed to the inhibition of AMP deaminase by Pi in the anoxic heart (Chen W, et al., 1996). We discuss the physiological significance of this heart-specific duality of degradation pathways.

Adenine↗
PubMed1996

A common pathway for p10 and calyx proteins in progressive stages of polyhedron envelope assembly in AcMNPV-infected Spodoptera frugiperda larvae.

The assembly of the polyhedron envelope in baculovirus-infected cells has been the subject of several studies, yet it is still poorly understood. We have used immunogold-labelled antibodies to two baculovirus proteins, p10 and calyx (also referred to as polyhedron envelope protein or PEP), to follow envelope assembly in AcMNPV-infected tissues of Spodoptera frugiperda larvae. We show that, in wild type virus, both proteins colocalize in fibrillar structures and associated electron-dense spacers which progress to encircle the polyhedra, as well as in completed polyhedron envelopes. In cells infected with polyhedrin-negative (PH-) viruses, an unusual proliferation of these spacers was observed suggesting a deregulatory event in the envelope assembly process. Results of Northern and Western blot analysis revealed that synthesis of P10 and calyx mRNA and proteins in PH- AcMNPV is unaffected as compared to wild type virus. Taken together, the observed physical and compositional connection between fibrillar structures, spacers and polyhedron envelopes, as well as the abnormal appearance of the spacers in PH- mutants, provide further evidence in support of a cooperative role of these structures in the assembly of the polyhedron envelope.

Animals↗
PubMed1996

Gingival fibromatosis.

Presented in this paper is a clinico-pathological report of 11 cases of gingival fibromatosis. Of the 11 patients in our series, 2 cases (18.2%) with family history began to have symptom from age of 3, which is manifested by the involvement of whole gum. 9 patients (81.8%), for which the etiology was unknown, had onset of the disease at the periods of deciduous dentition, mixed dentition or permanent dentition, with localized or overall involvement of gingiva. The disease is characterized by diffuse or localized proliferation of gum. Microscopically, the proliferating gum tissue revealed well-developed fibrotic structure with few cells; and the fibers were densely arranged without clear boundary. The fibrotic tissues showed a diminishing of blood vessels and the infiltration of inflammatory cells is not conspicuous.

Adolescent↗
PubMed1996

Identification of a group I intron in the nuclear small subunit rDNA of Pseudohalonectria lignicola.

A 865-bp insertion was detected within the nuclear small subunit (SSU) rDNA in two isolates (intron+) of the freshwater ascomycete Pseudohalonectria lignicola by the polymerase chain reaction (PCR). The intron sequences from the two isolates were identical to each other, and the exon sequences from the two intron+ isolates were identical to those in the intron- isolates in the PCR-amplified region of rDNA. Reverse transcription PCR (RT-PCR) indicated that the intron was absent in the mature rRNA. The intron sequence has all the characteristics of a group I intron, including four conserved sequence elements (P, Q, R, and S), the presence of a U at the 5' splice site of the exon, a G at the 3' splice site of the intron, a putative internal guiding sequence, and the sequence fit a secondary structure model for group I introns. Like most introns found in nuclear rDNA, this intron was located at a highly conserved region and was devoid of long open reading frames.

Ascomycota↗
PubMed1996

Characterization of subtypes of gamma-aminobutyric acid receptors in an Ascaris muscle preparation by binding assay and binding of PF1022A, a new anthelmintic, on the receptors.

We examined the effect of PF1022A, one of the gabergic anthelmintics newly developed in Japan, on gamma-aminobutyric acid (GABA) receptors using a radioligand binding technique in isolated membrane preparations of the nematode Ascaris suum. Membrane protein was prepared from the homogenate of somatic muscle cells after ultracentrifugation. In addition to the basic binding of [2,3-3H-(N)]-GABA, the radioligand [methyl-3H]-bicuculline is used to identify the GABAA receptor, whereas [butyl-4-3H]-baclofen is employed for GABAB receptor sites. The dissociation constants (Kd values) and the maximal numbers of binding sites (Bmax values) from Scatchard plotting for GABA receptors are close to those obtained in mammalian brain. PF1022A displaced in a concentration-dependent way the binding of [2,3-3H(N)]-GABA and [methyl-3H]-bicuculline as did other specific gabergic agents. In addition, PF1022A decreased the binding of [butyl-4-3H]-baclofen at a higher concentration, although this binding did not represent GABAB sites. In a comparison of the inhibition constants (Ki values) of PF1022A with those of other agents, it is conclusive that PF1022A bound with GABA receptors. A direct effect of PF1022A on GABA receptors can thus be postulated.

Animals↗
PubMed1996

Osteogenesis in extraskeletally implanted porous calcium phosphate ceramics: variability among different kinds of animals.

Synthetic porous calcium phosphate ceramics (HA/TCP) were implanted intramuscularly and subcutaneously in dogs, pigs, goats, rabbits and rats, designed to make a comparative study of the host tissue responses to porous HA/TCP ceramics in different kinds of animals. Specimens were harvested at 15, 30, 45, 60, 90 and 120 days after implantation. Decalcified and undecalcified sections were made and examined by light microscopy. Obvious bone formation could be detected in some specimens harvested from dogs and pigs after 45 days intramuscular implantation or after 60 days subcutaneous implantation. At days 90 and 120, an extensive amount of bone formed in all specimens implanted in dogs and pigs. However, no histologically detectable bone formation was observed in any specimen implanted intramuscularly and subcutaneously in goats, rabbits and rats until 120 days. It is demonstrated from this finding that the synthetic porous calcium phosphate ceramics are capable of inducing osteogenesis when implanted in non-bony sites, but this ability varies between different kinds of animals. Earlier periods of observation in specimens harvested from dogs showed that bone differentiation in the pore regions of the ceramics follows a complex process involving invasion of the fibrovascular connective tissues at day 15, appearance of polymorphic mesenchymal cells near the invading vasculature and at the interface with the ceramics at day 30, differentiation of osteoblasts and formation of bone matrix in direct contact with the surface of the ceramics at day 45, and finally remodelling of the fibrous connective tissue into an extensive amount of bone at days 60, 90 and 120.

Animals↗
PubMed1996

Electrophysiological and morphological properties of pyramidal and nonpyramidal neurons in the cat motor cortex in vitro.

Electrophysiological and morphological properties of the neurons in cat motor cortex were investigated using intracellular recording and staining techniques in a brain slice preparation. In response to intracellular injection of depolarizing current pulses, four distinct types of firing patterns were observed among cat neocortical neurons. Regular-spiking neurons were characterized by their repetitive firing from which conspicuous frequency adaptation was observed. Doublet-or-burst firing cells were marked with their tendency to fire 2-5 clustered spikes at the onset of depolarizing pulse. In doublet-or-burst firing neurons, but not in regular-spiking neurons, a low-threshold calcium current was revealed by single-electrode voltage clamp. Both regular-spiking and doublet-or-burst firing neurons had relatively wide action potentials. Fast-spiking neurons could fire extremely narrow action potentials at a very high frequency. Their frequency-to-intensity slope of steady-state firing was significantly higher than that of the other neurons. In contrast, narrow-spiking neurons had the smallest frequency-to-intensity slope for steady-state firing, although their action potentials were as narrow as those of the fast-spiking neurons. Both regular-spiking and doublet-or-burst firing neurons were identified as pyramidal neurons, and were found in all layers below layer I. Their apical dendrites were densely coated with dendritic spines. Narrow-spiking neurons were only recorded in layer V. They were large pyramidal cells with scare spines on their apical dendrites. Fast-spiking neurons were all nonpyramidal interneurons. Seven out of eight labelled fast-spiking cells had beaded dendrites without spines. Their axons had a large number of varicosities, and arborized extensively to form a dense plexus of terminals in the vicinity of their soma. The remaining neuron was found to be a spiny nonpyramidal neuron in layer V. These results demonstrate that, in addition to the three types of firing patterns previously identified in rodent neocortex, a group of neurons in the cat motor cortex express another type of firing behaviour which is characterized by extremely narrow action potential and very small frequency-to-intensity slope. Correlation with the morphological data shows that these neurons are large layer V pyramidal cells rather than nonpyramidal interneurons.

Action Potentials↗