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Biomedical subjects

C Tsai

Publications and source records attributed to C Tsai.

At least 37 records · Page 2Linked to original sources

Cloning, sequencing and tissue distribution of two related G protein-coupled receptor candidates expressed prominently in human lung tissue.

A novel G protein-coupled receptor, named GPR12A, was cloned by a PCR strategy using degenerate primers designed from sequences conserved among receptors for inflammatory mediators. Screening of a human lung cDNA library with GPR12A as a probe also identified a closely-related cDNA (GPR6C.1) that has been previously reported as GPR4 [13]. GPR12A and GPR6C.1 are 46.1% identical in amino acid sequence, but are less than 33% identical to any other known receptors. Northern analysis revealed that they are expressed prominently in the lung. Although the ligands for GPR12A and GPR6C.1 are unknown, their similarity suggests that they are receptors for ligands of similar or identical chemical nature.

Amino Acid Sequence↗

Biocompatibility of chemical-vapour-deposited diamond.

The biocompatibility of chemical-vapour-deposited (CVD) diamond surfaces has been assessed. Our results indicate that CVD diamond is as biocompatible as titanium (Ti) and 316 stainless steel (SS). First, the amount of adsorbed and 'denatured' fibrinogen on CVD diamond was very close to that of Ti and SS. Second, both in vitro and in vivo there appears to be less cellular adhesion and activation on the surface of CVD diamond surfaces compared to Ti and SS. This evident biocompatibility, coupled with the corrosion resistance and notable mechanical integrity of CVD diamond, suggests that diamond-coated surfaces may be highly desirable in a number of biomedical applications.

Absorption↗

Contraction potency of hypertrophic scar-derived fibroblasts in a connective tissue model: in vitro analysis of wound contraction.

Many investigators have reported that collagen gel contraction reflects the mechanism of wound contraction. By using a connective tissue model (CTM) of collagen gel lattice, we analyzed the contraction potency of fibroblasts that had been obtained from hypertrophic scar, normal skin, and normal oral mucosa. We then tried to analyze the mechanism of CTM contraction by immunofluorescent microscopic and transmission electron microscopic examinations. Hypertrophic scar-derived fibroblasts in CTM possessed the greatest contraction potency and the shortest lag time when compared with those of normal skin and normal oral mucosa-derived CTMs. It became clear that the initial contractive degree of CTM was closely related to morphological changes of fibroblast cells into bipolar and elongated shapes. Hypertrophic scar-derived fibroblasts elongated their processes faster, and their intracellular actin filaments were more numerous than those of normal fibroblasts. We believe that the hypertrophic scar-derived CTM is a useful pathological model for the research of wound contraction and hypertrophic scar formation.

Adolescent↗

Pair-rule expression of the Drosophila fushi tarazu gene: a nuclear receptor response element mediates the opposing regulatory effects of runt and hairy.

The segmentation genes runt and hairy are required for the proper transcriptional regulation of the pair-rule gene fushi tarazu during the blastoderm stage of Drosophila embryogenesis. The expression of different fushi tarazu reporter genes was examined in runt and hairy mutant embryos, as well as in runt over-expressing embryos in order to identify DNA elements responsible for mediating these regulatory effects. The results indicated that runt and hairy act through a common 32 base-pair element. This element, designated as fDE1, contains a binding site for a small family of orphan nuclear receptor proteins that are uniformly expressed in blastoderm embryos. The pair-rule expression of reporter gene constructs containing multimerized fDE1 elements depends on activation by runt and repression by hairy. Examination of reporter genes with mutated fDE1 elements provided further evidence that this element mediates both transcriptional activation and repression. Genetic experiments indicated that the opposing effects of runt and hairy were not due solely to cross-regulatory interactions between these two genes and that fDE1-dependent expression is regulated by factors in addition to runt and hairy.

Animals↗

Expression of CD60 on multiple cell lineages in inflammatory synovitis.

BACKGROUND: CD60 is a recently described T cell subset marker that is expressed on the surface of most T lymphocytes in synovial tissue and fluid and on a smaller proportion of peripheral T cells. Activation of T lymphocytes can be triggered through CD60. CD60 is also expressed by neuroectodermally derived cells in thymic epithelium and in skin. EXPERIMENTAL DESIGN: Immunohistologic analysis of CD60 expression in synovium and thymus was performed using formalin-fixed tissue samples. Nonlymphoid cell lines grown from similar tissues were analyzed by flow cytometry. RESULTS: CD60 was readily identified in formalin-fixed, paraffin-embedded tissues. Simultaneous examination of CD60 distribution and cell morphology demonstrated that, in addition to its presence on T cells, CD60 was also expressed by a variety of nonlymphoid cells in synovium, including synovial lining cells, vascular endothelium, and dendritic-appearing cells deep within synovial tissue. Synovial tissue expression of CD60 was similar in rheumatoid arthritis and in other forms of inflammatory arthritis. In addition, it was strongly expressed by giant cells in pigmented villonodular synovitis. Surface expression of CD60 was detected by flow cytometry on cultured synoviocytes and on other CD60+ nonlymphoid cells, thus excluding adsorption of CD60 shed by T cells as a sufficient explanation of the immunohistologic findings. CONCLUSIONS: These results define the T cell-activating CD60 determinant as a broadly distributed Ag within synovial tissue, with a possible functional role in the activation of a variety of cellular populations. CD60 may also be a marker for previously undescribed cell subsets in the synovial compartment, possibly including a cell population of neuroectodermal origin.

Adult↗

Gap gene properties of the pair-rule gene runt during Drosophila segmentation.

The Drosophila Runt protein is a member of a new family of transcriptional regulators that have important roles in processes extending from pattern formation in insect embryos to leukemogenesis in humans. We used ectopic expression to investigate runt's function in the pathway of Drosophila segmentation. Transient over-expression of runt under the control of a Drosophila heat-shock promoter caused stripe-specific defects in the expression patterns of the pair-rule genes hairy and even-skipped but had a more uniform effect on the secondary pair-rule gene fushi tarazu. Surprisingly, the expression of the gap segmentation genes, which are upstream of runt in the segmentation hierarchy was also altered in hs/runt embryos. A subset of these effects were interpreted as due to an antagonistic effect of runt on transcriptional activation by the maternal morphogen bicoid. In support of this, expression of synthetic reporter gene constructs containing oligomerized binding sites for the Bicoid protein was reduced in hs/runt embryos. Finally, genetic experiments demonstrated that regulation of gap gene expression by runt is a normal component of the regulatory program that generates the segmented body pattern of the Drosophila embryo.

Animals↗

Scleroderma after silicone augmentation mammoplasty: report of a case.

The development of connective tissue diseases in patients who have had silicone gel breast implants has not been previously described in Taiwan. Herein, we report a 44-year-old female patient who underwent a silicone gel breast implant approximately 10 years prior to the onset of scleroderma. She developed generalized sclerosis, severe defective esophageal clearance, and severe reduction of gas exchange with DLco 37% of predicted value. Immunologic tests revealed that the antinuclear antibody titer was 1:2,560 with speckled pattern and positive anti-Scl-70 antibody. In spite of prednisolone and D-penicillamine treatment, her condition deteriorated. However, following the removal of the prosthesis, short-term subjective and objective improvements in both skin lesions and pulmonary function were noted. It is important that plastic surgeons, rheumatologists, and dermatologists, as well as primary care physicians, be aware of the possible association of sclerosis with silicone gel breast implants.

Adult↗

An anti-CD2 monoclonal antibody that both inhibits and stimulates T cell activation recognizes a subregion of CD2 distinct from known ligand-binding sites.

The T lymphocyte glycoprotein CD2 appears to have an important role in human T cell development and activation. A novel anti-CD2 monoclonal antibody, designed UMCD2, was shown to block E rosetting, and therefore was defined as recognizing the Tll1 ligand-binding epitope. Binding of UMCD2 to T cells and thymocytes was blocked by several, but not all, anti-Tll1 antibodies, suggesting that the Tll1 epitope consists of more than one subepitope. In functional studies, the combination of UMCD2 plus anti-Tll3 was mitogenic for T cells; in some individuals, the level of activation was as high as that seen for the combination of anti-Tll2 plus anti-Tll3. However, when UMCD2 was added to other stimuli mitogenic for T lymphocytes, such as IL-2 or anti-CD3-Sepharose, it inhibited T cell responses. Although the combination of UMCD2 and anti-Tll3 induced an increase in cytoplasmic free calcium, the inhibitory activities of UMCD2 were not accompanied by effects on calcium fluxes. A panel of previously characterized CD2 mutants was then analyzed for binding of UMCD2 and other anti-CD2 monoclonals. Surprisingly, UMCD2 bound to all mutants tested, although the other anti-CD2 antibodies with specificity for the ligand-binding region of CD2 each failed to bind to one or more mutants. These data suggest that binding of antibody to a particular CD2 epitope can have opposite effects on the state of T cell activation, depending on the costimulus. Moreover, inhibitory effects mediated through CD2 may use a signaling mechanism distinct from that used in CD2 pathway activation. Of particular interest, the portion of the CD2 ligand-binding region recognized by UMCD2 is distinct from areas of the CD2 molecule that have previously been studied.

Amino Acid Sequence↗

Glycation mediated lens crystallin aggregation and cross-linking by various sugars and sugar phosphates in vitro.

Glycation of lens crystallins results in protein conformational changes, oxidation, browning and aggregation. Though glucose is the major sugar, other sugars and sugar phosphates generated as intermediates of metabolic pathways are present in the lens, albeit at low concentrations. In this study we incubated bovine lens soluble fraction with various sugars and sugar phosphates (5mM for 10 days). The reactivity was in the order trioses > tetroses > pentoses > hexoses. High molecular weight (HMW) aggregates were also formed at a comparable rate. Increased levels of fluorescence were associated with the HMW aggregates with fast reacting sugars. The phosphorylated derivatives were only slightly more reactive than their respective sugars. Interestingly, fructose-1,6-diphosphate was more reactive and cross-linked more readily than fructose-6-phosphate. Gel electrophoresis under reducing and nonreducing conditions showed formation of disulfide linked protein aggregates with slow reacting sugars such as glucose and non-disulfide covalent linked protein aggregates with fast reacting sugars such as erythrose. In contrast, if 0.1 m DTT was present in erythrose incubations (a fast reacting sugar), the HMW aggregate formation was significantly reduced. In order to show the reactivity among the slow reacting hexoses, we incubated lens proteins with 1 M hexoses for 30 days and the results showed that galactose was more reactive and showed higher cross-linking than fructose and glucose. These results thus indicate that relatively low levels of some sugars and sugar phosphates in the lens could be compensated by enhanced lens protein cross-linking and the combined effect could be rather significant with respect to cataractogenesis.

Animals↗

Retention of retinal axon collateral is responsible for induced ipsilateral retinotectal projections in adult goldfish.

In normal goldfish, optic axons innervate only the contralateral optic tectum. When one eye was enucleated and the optic nerve of the other eye crushed, the regenerating optic axons innervated both optic tecta. We studied the presence of bilaterally projecting retinal ganglion cells by double retrograde cell labeling methods using Nuclear Yellow and True Blue dyes. About 10% of the retinal ganglion cells were double labeled and these cells were found throughout the retina. In addition, HRP application to the ipsilateral tectum revealed retrogradely-labeled retinal ganglion cells of all morphological types. These results suggest that induced ipsilateral projections are formed by regenerating axon collaterals and that all cell types are involved in the generation of normal mirror image typography.

Animals↗

Enhancement of the antiviral activity of poly r(A-U) by ametantrone and mitoxantrone.

The role of ametantrone (HAQ) and mitoxantrone (DHAQ) in modulating the antiviral and interferon-inducing activities of poly r(A-U) was examined using the human foreskin fibroblast-vesicular stomatitis virus (HSF-VSV) bioassay system in which the concentration of poly r(A-U) was fixed at 0.05 mM or 0.2 mM while the HAQ or DHAQ concentration was varied to produce variable HAQ (or DHAQ)/ribonucleotide ratios ranging from 1:16 to 2:1. HAQ, DHAQ and poly r(A-U) tested individually were not efficacious antiviral agents. When poly r(A-U) was combined with the ametantrone or mitoxantrone the antiviral activity was potentiated 10-fold at HAQ (or DHAQ)/ribonucleotide ratios in the region of 1/4 to 1/6. The interferon-inducing activity of the HAQ (or DHAQ)/poly r(A-U) combinations were equal to the sum of the interferon-inducing activity of the poly r(A-U) and the HAQ (or DHAQ). These results indicate that the HAQ and DHAQ potentiate the antiviral activity of the poly r(A-U) without the superinduction of interferon. The direct viral inactivation study demonstrated that HAQ, DHAQ, poly r(A-U) and the HAQ (or DHAQ)/poly r(A-U) combinations did not inactivate the VSV at concentrations near the viral 50% inhibitory dose.

Biological Assay↗

Complex modes of heat shock factor activation.

Eucaryotic organisms respond to elevated environmental temperatures by rapidly activating the expression of heat shock genes. The transcriptional activation of heat shock genes is mediated by a conserved upstream regulatory sequence, the heat shock element (HSE). Using an HSE-binding assay, we show that a cellular factor present in a range of vertebrate species binds specifically to the HSE. This factor is presumably the transcriptional activator of heat shock genes, heat shock factor (HSF). In vertebrates, the binding of HSF to the HSE was induced when cells were subjected to heat shock at high temperatures, even in the absence of protein synthesis. Under mild heat shock conditions, HSF binding was induced to a lesser extent, but this induction required protein synthesis, suggesting that synthesis of HSF itself, or an activating factor, is necessary for response to heat shock at intermediate temperatures. The inducibility of HSF binding in higher eucaryotes is contrasted with constitutive HSF binding activity in fungi. It appears that despite conservation of the HSE in evolution, the means by which HSF is activated to bind DNA in higher and lower eucaryotes may have diverged.

Animals↗

Racial differences in optic nerve head parameters.

Results of previous studies have strongly indicated that the prevalence of elevated intraocular pressure is greater in blacks than in whites and that blacks are more susceptible than whites to glaucomatous damage at any given level of pressure. It has also been suggested that a larger disc area might predispose an eye to glaucomatous damage. We investigated the possibility that clinically quantifiable differences might exist in optic disc parameters between normotensive white and black patients. Disc area, cup-to-disc ratio, and cup volume measured with a video-ophthalmograph (Rodenstock Optic Disc Analyzer) were significantly larger in blacks than in whites, while there was no difference in the disc rim area between the two groups. We derived a mathematical model of the optic disc that relates posterior displacement of the lamina cribrosa to the disc area, distensibility of the disc, and intraocular pressure.

Adolescent↗

Endocytosis in the rat retinal pigment epithelium.

Endocytosis in the retinal pigment epithelium (RPE) of rats was studied using horseradish peroxidase, microperoxidase and ferritin tracers. Tracer uptake was mediated by coated pits and coated vesicles. Coated pits formed at two discrete regions at the RPE plasma membrane: that portion of basal membrane directly opposing Bruch's membrane, and at the bases of the apical lamellae and villi. Two populations of coated vesicles were identified and distinguished by size, location and function. Large coated vesicles (91.8 +/- 14.7 nm in diameter) were located near the cell surface and incorporated tracer. Small coated vesicles (64.5 +/- 15.7 nm diameter) located more deeply within the cell were not tracer-labeled, and were often fused with the endoplasmic reticulum or the Golgi apparatus. Observations of the endocytic pathway in rat RPE cells are presented. Tracer was also found in organelles of the lysosomal system, e.g. the multivesicular body, but was not identified in the smooth endoplasmic reticulum or Golgi apparatus.

Animals↗

Trans-2-carboxy-3-pyrrolidineacetic acid (CPAA), a novel agonist at NMDA-type receptors.

Although 2-carboxy-3-pyrrolidineacetic acid (CPAA) analogs are associated with activity as agonists at kainate-type receptors, here we report that CPAA is an agonist at N-methyl-D-aspartic acid-type receptors. CPAA evoked the release of [3H]acetylcholine (ACh) from striatal slices with the same efficacy as NMDA, and an EC50 of 20.0 microM, compared to an EC50 of 45.8 microM for NMDA. CPAA-evoked [3H]ACh release was inhibited by CPP (IC50 = 5.1 microM), tiletamine (IC50 = 0.53 microM), MK-801 (IC50 = 0.12 microM), and MgCl2 (IC50 = 26 microM). CPAA produced a tachyphylaxis when applied continuously for 18 min or more, and a cross-tachyphylaxis to NMDA. Similarly, NMDA generated a cross-tachyphylaxis to CPAA. All of these data suggest that CPAA is an agonist at NMDA-type receptors.

Acetylcholine↗

CGS 19755 is a potent and competitive antagonist at NMDA-type receptors.

The N-Methyl-D-aspartate (NMDA)-type receptor blocking properties of CGS 19755, a novel, rigid analog of 2-amino-5-phosphonopentanoate, were demonstrated in vitro by the ability of the compound to block NMDA-evoked [3H]acetylcholine release (pA2 = 5.93). CGS 19755 (0.045 and 0.224 mmol/kg i.p.) was shown to be active in vivo as well by its ability to block harmaline-induced increases in cerebellar cGMP. Finally, CGS 19755 blocked sound-induced seizures in DBA/2 mice completely at doses of 1.0 nmol i.c.v. or 0.1 mmol/kg i.p. Taken together, these data indicate that CGS 19755 is a potent and competitive NMDA antagonist in vitro which is also active in vivo.

Acetylcholine↗

Glycine-evoked release of [3H]acetylcholine from rat striatal slices is independent of the NMDA receptor.

KCl-, NMDA-, and glycine-evoked release of [3H]acetylcholine was studied in superfused rat striatal slices. KCl-evoked release of [3H]acetylcholine was inhibited by 1.2 mM MgCl2 and 100 microM lidocaine. Similarly, NMDA-evoked release was inhibited by MgCl2 and lidocaine as well as 10 microM CGS 19755, a competitive antagonist at NMDA receptors, and 10 nM MK-801, a noncompetitive antagonist of NMDA-induced responses. Glycine-evoked release was calcium-dependent and was inhibited by 0.1 microM strychnine whereas KCl- and NMDA-evoked release were resistant to strychnine. In addition, lidocaine inhibited the glycine-induced response. Cross-tachyphylaxis was not observed between NMDA- and glycine-evoked release. These results indicate that the strychnine-sensitive, glycine-evoked release of [3H]acetylcholine is independent of the NMDA receptor.

Acetylcholine↗

Enhancement of the antiviral and interferon-inducing activities of poly r(A-U) by carminic acid.

Experiments have been designed to systematically examine the effects of carminic acid (CAR) on the antiviral/interferon-inducing activity of poly r(A-U), using the human foreskin fibroblast-vesicular stomatitis virus bioassay system. Modulation of the antiviral/interferon-inducing activity of poly r(A-U) by carminic acid was examined at fixed poly r(A-U) concentrations of 0.05 mM or 0.2 mM while varying the carminic acid concentrations to produce variable CAR/ribonucleotide ratios ranging from 1:16 to 2:1. Carminic acid and poly r(A-U) were tested individually at the concentrations employed in the CAR/poly r(A-U) combinations. Neither the carminic acid alone nor poly r(A-U) alone were effective antiviral agents/interferon inducers. The antiviral/interferon-inducing activity of poly r(A-U) was potentiated twelve-fold at CAR/ribonucleotide ratios in the region of 1/6 to 1/4. These results suggest a synergism between the poly r(A-U) and the carminic acid at the concentrations employed in this study.

Anthraquinones↗