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Biomedical subjects

S Chang

Publications and source records attributed to S Chang.

At least 289 records · Page 16Linked to original sources
PubMed1991

Efficient immortalization of luminal epithelial cells from human mammary gland by introduction of simian virus 40 large tumor antigen with a recombinant retrovirus.

When defined in terms of markers for normal cell lineages, most invasive breast cancer cells correspond to the phenotype of the common luminal epithelial cell found in the terminal ductal lobular units. Luminal epithelial cells cultured from milk, which have limited proliferative potential, have now been immortalized by introducing the gene encoding simian virus 40 large tumor (T) antigen. Infection with a recombinant retrovirus proved to be 50-100 times more efficient than calcium phosphate transfection, and of the 17 cell lines isolated, only 5 passed through a crisis period as characterized by cessation of growth. When characterized by immunohistochemical staining with monoclonal antibodies, 14 lines showed features of luminal epithelial cells and of these, 7 resembled the common luminal epithelial cell type in the profile of keratins expressed. These cells express keratins 7, 8, 18, and 19 homogeneously and do not express keratin 14 or vimentin; a polymorphic epithelial mucin produced in vivo by luminal cells is expressed heterogeneously and the pattern of fibronectin staining is punctate. Although the cell lines have a reduced requirement for added growth factors, they do not grow in agar or produce tumors in the nude mouse. When the v-Ha-ras oncogene was introduced into two of the cell lines by using a recombinant retrovirus, most of the selected clones senesced, but one entered crisis and emerged after 3 months as a tumorigenic cell line.

Animals↗
PubMed1991

Molecular characterization of erythrocyte glycophorin C variants.

Human erythrocyte glycophorin C plays a functionally important role in maintaining erythrocyte shape and regulating membrane mechanical stability. Immunochemical and serologic studies have identified a number of glycophorin C variants that include the Yus, Gerbich, and Webb phenotypes. We report here the molecular characterization of these variants. Amplification of glycophorin C mRNA from the Yus phenotype, using two oligonucleotide primers that span the coding domain, generated a 338-bp fragment compared with a 395-bp fragment generated by amplification of normal glycophorin C mRNA. Sequencing of the mutant 338-bp fragment identified a 57-bp deletion that corresponds to exon 2 of the glycophorin C gene. Similar analysis showed deletion of 84-bp exon 3 in the Gerbich phenotype. In contrast to the generation of shorter than normal DNA fragments from mRNA amplification in the Yus and Gerbich phenotypes, amplification of mRNA from the Webb phenotype generated a normal-sized fragment. Sequencing of this DNA fragment showed an A----G substitution at nucleotide 23 of the coding sequence, resulting in the substitution of asparagine by serine. This modification accounts for the altered glycosylation of glycophorin C seen in this phenotype. These results have enabled us to characterize glycophorin C variants in three different phenotypes that involve deletions of exons 2 and 3 of the glycophorin C gene, as well as a point mutation in exon 1 that results in altered glycosylation of this protein.

Amino Acid Sequence↗
PubMed1991

Are some chondrule rims formed by impact processes? Observations and experiments.

Observations and experimental evidence are presented to support the hypothesis that high-speed impact into a parent body regolith can best explain certain textures and compositions observed for rims on some chondrules. A study of 19 interclastic rimmed chondrules in the Weston (H 3/4) ordinary chondrite shows that two main rim types are present on porphyritic olivine-pyroxene (POP) and porphyritic pyroxene (PP) chondrules: granular and opaque rims. Granular rims are composed of welded, fine-grained host chondrule fragments. Bulk compositions of granular rims vary among chondrules, but each rim is compositionally dependent on that of the host chondrule. Opaque rims contain mineral and glass compositions distinctly different from those of the host, partially reacted chondrule mantle components, and some matrix grains. Opaque rims are greatly enriched in FeO (up to 63 wt%). The original chondrule pyroxene compositional zonation patterns and euhedral grain outlines are discontinuous at the chondrule/rim interface. Opaque rims are dominated by fayalitic olivine (Fa92-56), with high Al2O3 content (0.78-3.15%), which makes them distinctly different from primary olivine, but similar to Fe-olivine in chondrule rims of other meteorites. Thin zones of chondrule minerals adjacent to the present rims are intermediate in FeO content between the Mg-rich interior and the Fe-rich rim, which indicates a reaction relationship. Regardless of conclusions drawn regarding other types of rims, granular and opaque rim characteristics appear to be inconsistent with nebular condensation, in that host and matrix fragments are included within the rim. We have initiated a series of experiments, using the Ames two-stage light gas gun, to investigate the hypothesis that the Weston chondrule rims are the result of thermal and mechanical alteration upon impact into a low-density medium. Clusters of approximately 200-micron-sized silicate particles were fired into aerogel (density = 0.1 g cm-3) at velocities of 5.6, 4.7, and 2.2 km sec-1. Recovered grains show characteristics that range from fragmented projectile grains mixed with melted aerogel that nearly rim the grains to grains that have melted aerogel clumps mixed with partially melted projectile. These experimental results demonstrate that rim-like thermal and mechanical alteration of projectiles can result from a high-velocity encounter with a low-density target. Therefore, experiments using appropriately chosen projectile and target materials can provide a test of the hypothesis that chondrule rims common to Weston and possibly other ordinary chondrites were formed by such a process.

Astronomical Phenomena↗
PubMed1991

A chemiluminescent immunoassay for urinary thromboxane B2.

Because of the vasoactive properties of thromboxane A2 and other related prostaglandins, much research has been conducted on drugs which alter their levels. Urinary levels of thromboxane B2 and 2,3-dinor thromboxane B2 (major urinary metabolite of thromboxane B2) are used as an indication of thromboxane production in-vivo. In order to accurately measure urinary TXB2 levels of subjects on investigative drugs which lower TXA2 and subsequently TXB2, a simple and sensitive analytical tool becomes necessary. We have thus developed a non-radioisotopic (chemiluminescent) assay for urinary TXB2. Sensitivity has been demonstrated to 5 pg/ml. The method correlates well with gas chromatography/mass spectrometry (the accepted reference method) even without column chromatographic purification prior to the conduct of the chemiluminescent assay (r = 0.96). In addition, we have demonstrated feasibility for a chemiluminescent assay to measure urinary 2,3-dinor TXB2.

Acridines↗
PubMed1991

Characterization of acetylcholinesterase molecular forms of the root-knot nematode, Meloidogyne.

Multiple molecular forms of acetylcholinesterase have been isolated and characterized from the root-knot nematodes Meloidogyne arenaria and Meloidogyne incognita. The forms of enzyme present in these 2 species are similar but not identical to those that occur in the free-living nematode Caenorhabditis elegans. The 5 enzyme forms exhibit differential solubilities and can be classified into 3 classes, A, B, and C, based on substrate affinity, inhibitor and detergent sensitivity, and thermal inactivation profiles. An unusual class of acetylcholinesterase has been isolated from Meloidogyne which has very high affinity for acetylcholine, but is highly resistant to carbamate and organophosphate inhibitors. The potential roles of the molecular forms in nematode behavior and sensitivity to nematicides are discussed.

Acetylcholinesterase↗
PubMed1991

DM-GRASP, a novel immunoglobulin superfamily axonal surface protein that supports neurite extension.

We have identified a 95 kd cell surface protein, DM-GRASP, that is expressed on a restricted population of axons. Its expression begins early in chick embryogenesis, and within the spinal cord it is localized to axons in the dorsal funiculus, midline floorplate cells, and motoneurons. Antibodies to DM-GRASP impair neurite extension on axons, and purified DM-GRASP supports neurite extension from chick sensory neurons. We have cloned and sequenced the cDNA corresponding to this protein and find that it is a new member of the immunoglobulin superfamily of adhesion molecules. Consequently we have named this protein DM-GRASP, since it is an immunoglobulin-like restricted axonal surface protein that is expressed in the dorsal funiculus and ventral midline of the chick spinal cord.

Activated-Leukocyte Cell Adhesion Molecule↗
PubMed1991

Translocation of the nuclear autoantigen La to cell surface: assembly and disassembly with the extracellular matrix.

La (SS-B) protein is known as one major antigenic target for autoantibodies from patients with certain autoimmune diseases such as Sjogren's syndrome or Lupus Erythematosus. La protein belongs to the so called "extractable nuclear antigens". Here we report that La antigen is not restricted to the nucleus as one might deduce from the exclusive nuclear staining pattern of patient anti-La antibodies but after stimulation of serum-starved cells with 10% fetal calf serum (FCS) appears and stays for at least 45 min at the outer surface of CV-1 cells being available for binding of anti-La antibodies. In addition we found that a minor part of La antigen associates with the extracellular fibronectin network. After addition of 10% FCS to serum starved cells this extracellular autoantigen disassembled from the extracellular matrix and was taken up again by the cells. Incubation of serum starved cells with mercuric chloride, a known potent inducer of autoantibodies, also resulted in a detachment of the extracellular matrix associated La protein. From our studies it becomes likely that La protein itself is the antigen during autoimmunization. Moreover, once developed, anti-La antibodies might be able to bind to cell surface expressed La protein resulting in a damage of these cells leading to the inflammational events known to occur during disease.

Autoantigens↗
PubMed1991

Perfluorocarbon liquids in the management of complicated retinal detachments.

Low viscosity perfluorocarbon liquids have unique physical properties making them ideal for the temporary tamponade, mechanical fixation, and manipulation of the retina. These substances are optically clear, have a specific gravity greater than that of water, and have interfacial tension properties similar to those of silicone oil. The temporary mechanical fixation of the retina by perfluorocarbon liquids facilitates the removal of epiretinal membranes and release of tractional forces. Intraoperative flattening of the retina under perfluorocarbon liquid eliminates the need for a posterior retinotomy. Perfluorocarbon liquids are being used intraoperatively for hydrokinetic manipulation of the retina during vitrectomy for complicated retinal detachments secondary to proliferative vitreoretinopathy, giant retinal tears, and trauma. Additional indications for the intraoperative utilization of perfluorocarbon liquids in vitreous surgery are discussed.

Eye Diseases↗
PubMed1991

Aneurysm arising from the branch of the superior mesenteric artery.

Superior mesenteric branch aneurysms are rare and usually become symptomatic at the time of rupture. Pain, gastrointestinal blood loss and intra-abdominal hemorrhage draw attention to the presence of aneurysms in 70% of the reported cases. We report on a 64-year-old male patient who had an emergent laparotomy for acute abdominal pain at a local hospital in Changhua in March of 1988. The operative finding was an unresectable mesentric mass, and the pathologic finding of the biopsy was a chronic abscess. Because of two episodes of tarry stools after the operation, the patient was referred to the National Taiwan University Hospital for further evaluation of the intra-abdominal mass. After admission in April of 1990, abdominal sonogram and CT examinations demonstrated the presence of a multilobulated mass which was suspected to be an aneurysm. Selective superior mesenteric arteriography confirmed this diagnosis and showed that the aneurysm arose from the origin of the ileocolic branch. At surgery, the aneurysm was found to have a fistula tract communicating with the terminal ileum. The aneurysm and the associated segment of the terminal ileum were successfully removed. We herein report this unusual case.

Aneurysm↗
PubMed1991

Experimental studies of tolerance to intravitreal perfluoro-n-octane liquid.

Low viscosity perfluorocarbon liquids are useful adjuncts during vitreoretinal surgery. This study evaluates perfluoro-n-octane (PFnO) for intraoperative use by examining ocular tolerance in rabbits and pigs. Rabbits were treated with intravitreal placement of PFnO for 48 hours and were examined during a period of 2 months following treatment. No clinical, electroretinographic, or light and electron microscopic evidence of adverse effects on the retina were observed. Small residual amounts of PFnO produced no adverse histologic changes 6 months after treatment. In pigs treated with mechanical vitrectomy and placement of PFnO for 3 hours, electron microscopic evaluation of the retina confirmed that the liquid had been well tolerated. These findings indicate that PFnO is safe for intraoperative use. When PFnO remained in the vitreous cavity of rabbits for longer than 1 week, varying degrees of globule dispersion occurred. Furthermore, histologic examination revealed changes in the inferior retina: photoreceptor outer segments were distorted, the outer plexiform layer was narrowed, and preretinal accumulation of macrophages had occurred.

Animals↗
PubMed1990

Characterization of putative cytoskeletal proteins from a trypanosomatid and their comparative binding to microtubules and soluble tubulin.

The corset of microtubules which encloses the cell body of Crithidia fasiculata displays cross-links joining tubules to each other and to plasma membrane. Two proteins, designated COP-33 and COP-61 on the basis of their subunit Mr values, have been considered putative components of this apparatus because of their abundance in isolated cytoskeleton and their ability to cross-link brain microtubules in vitro. The oligomeric structures of the native proteins have now been characterized, and they have been shown to be basic, rather symmetrical, and to require detergent for solubilization. Using monospecific antibodies, enzyme-linked immunoassays of subcellular fractions have shown that each is 5-fold enriched in the microtubule plasma membrane fraction and absent from flagellar and some internal membranes. The binding of each to soluble tubulin and to microtubules has been systematically studied and compared with that of two noncytoskeletal protein ligands (glycolytic enzymes). The observed positive cooperatively was unexpected for binding of these large ligands to microtubules. In each case the maximum number of mol of ligand bound per mol of tubulin (0.5-1.0) was identical whether the latter was dissociated or assembled and so were the dissociation constants (1.3-6.0 x 10(-7) M) for three of the ligand proteins.

Alkaloids↗
PubMed1990

Transgenic mice with intracellular immunity to influenza virus.

We have generated transgenic mice that express the intracellular anti-influenza virus protein Mx1 under control of an interferon-responsive regulatory element. Upon infection with influenza virus, mice of a high responder line produce Mx1 protein locally at the sites of initial viral replication, exhibit little viral spread, and survive infection. Mice of a low responder line show more extensive viral spread and survive infection only when virus is given at high doses. To survive low dose infections, these mice require injection of interferon along with virus. The results show that influenza viral pathogenesis is determined by a subtle balance between the dose of the infecting virus and the levels of the antiviral host factor Mx1 and that mice can be rendered resistant to a virulent infection by "intracellular immunization" achieved through germline transformation.

Animals↗
PubMed1990

SLACS retrotransposon from Trypanosoma brucei gambiense is similar to mammalian LINEs.

We have characterized a retrotransposon in Trypanosoma brucei gambiense uniquely associated with the spliced-leader (SL) RNA gene cluster (Spliced Leader Associated Conserved Sequence, SLACS). There are nine copies of SLACS and DNA sequence analysis of one shows the hallmarks of Line-1 like elements. SLACS has generated a 49 bp target DNA duplication at its insertion site and its 3'-end is preceded by a poly(A) stretch. Two putative open reading frames (ORFs) span 75% of the element. ORF1 has CysHis motif associated with the retroviral gag polypeptide while ORF2 shows homology with reverse transcriptase sequences. Its 5'-end contains a repeated segment of a 185 bp that varies in copy number in different SLACS insertions. Retrotransposon-like sequences inserted into the SL-RNA genes occur in several hemoflagellates. These elements may represent a related family which has maintained its target site specificity.

Amino Acid Sequence↗