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Biomedical subjects

C M Chen

Publications and source records attributed to C M Chen.

At least 253 records · Page 14Linked to original sources
PubMed1986

Analysis of the expression of H-2K, D and L locus mRNAs by use of an oligonucleotide primer.

By employing an oligonucleotide-dependent cDNA synthesis/HaeIII digestion procedure followed by gel electrophoretic separation of HaeIII digested cDNAs, we have developed a method for analyzing the expression of H-2 mRNAs encoding the K, D and L locus histocompatibility antigens. This method provides unambiguous data as the individual K, D or L specific cDNA bands can be isolated from the gel and partially sequenced. The method is readily applicable to analyzing the relative expression of K, D and L locus mRNAs in normal mouse tissues and their tumors.

Animals↗
PubMed1986

Nucleotide sequence of the structural genes for an anion pump. The plasmid-encoded arsenical resistance operon.

The structural genes for the arsenical pump of the conjugative R-factor R773 contained on a HindIII fragment of 4.3 kilobase pairs were cloned into bacteriophage M13. A series of ordered deletions was created using Bal31 digestion, and the nucleotide sequence of the operon determined. Three open reading frames for genes arsA, arsB, and arsC were found. The arsA gene encodes a hydrophilic protein of 63,169 Da with two potential adenylate-binding sites. The arsB gene encodes a potentially membrane protein of 45,577 Da. The arsC gene encodes a 15,811-Da hydrophilic protein. The arsA and arsC gene products correspond to cytosolic proteins previously identified from minicell experiments. Isolated ArsA protein was shown to bind to dye-agarose columns which act as affinity resins for nucleotide-binding proteins. A model is proposed in which these gene products form an anion translocating ATPase for extrusion of arsenite and arsenate from resistant cells.

Adenosine Triphosphatases↗
PubMed1986

Degradation kinetics and mechanism of N6-[(dimethylamino)methylene]mitomycin C in aqueous solutions.

The degradation of N6-[(dimethylamino)methylene]mitomycin C, a semisynthetic analogue of mitomycin C, was studied in aqueous solution. The compound degraded rapidly and followed pseudo-first-order kinetics in both acidic (pH less than 5) and basic pH greater than or equal to 9) media. In the near-neutral pH region, however, biphasic kinetics were observed. At the pH of maximum stability (6.5), 10% activity was lost after approximately 6 h at 22 degrees C. Citrate and phosphate species were catalytic at pH 6.5. Spectrophotometric and HPLC methods were used to elucidate the degradation mechanism at pH 7-9. Under these conditions, equilibrium addition of one water molecule into the amidine side chain occurred, followed by parallel formation of mitomycin C and N6-(formyl)mitomycin C. The latter compound subsequently hydrolyzed to mitomycin C.

Buffers↗
PubMed1986

Confirmation of human cytomegalovirus by reverse passive hemagglutination with monoclonal antibodies reactive to the major glycosylated peptide (GP-66).

Sheep erythrocytes coated with three monoclonal antibodies, each reactive to a different epitope of the 66-kilodalton cytomegalovirus (CMV) matrix protein, were used in a reverse passive hemagglutination test with CMV-infected cell lysate to identify and confirm the CMV. The test is specific only for CMV, since 5 laboratory strains of CMV (AD169, Davis, Espilat, C-87, and Towne) and 10 clinical isolates reacted well, but uninfected MRC-5 cell lysate, lysates of herpes simplex virus types 1 and 2, varicella-zoster virus, and adenoviruses did not react. The reactive CMV lysate was confirmed by the pretreatment of CMV lysate with the three monoclonal antibodies followed by the addition of antibody-coated erythrocytes. The reverse passive hemagglutination test and the confirmatory blocking test are performed at the same time, requiring 2 h to complete. Since V-bottom microtiter 96-well plates and a 25-microliter pipette can be used to perform the test, it is ideal for CMV confirmation, especially when the equipment to read the fluorescent-antibody test or enzyme-linked immunosorbent assay is not available.

Antibodies, Monoclonal↗
PubMed1985

Percutaneous transsplenic balloon dilatation for stenotic distal splenorenal shunt.

A 27 year old cirrhotic male underwent stenosis of the distal splenorenal shunt to treat bleeding of esophageal varices. Percutaneous transluminal angioplasty was performed first by catheterization of the right femoral vein, but without success. Therefore, a direct puncture of the splenic vein through the spleen was made, under sonographic guidance, followed by balloon dilatation of the stenotic anastomosis. The pressure of the splenic vein before and after PTA was 35 and 29 cm H2O, respectively. Both the follow-up barium esophagogram and esophagoscopy showed marked regression of the esophageal varices.

Adult↗
PubMed1985

Cytokinin-modulated gene expression in excised pumpkin cotyledons.

Comparison of two-dimensional polyacrylamide gel electrophoretic maps of proteins isolated from benzyladenine-treated and untreated pumpkin (Cucurbita pepo L. cv Halloween) cotyledons showed that the expression of certain proteins is enhanced, induced, or suppressed by the cytokinin treatment. The amount of poly(A)(+) mRNA isolated from cotyledons incubated with 10(-4) molar benzyladenine for five days was about four-fold over the water-incubated control. The activity of hydroxypyruvate reductase prepared from purified cotyledonous microbodies and analyzed by native gel electrophoresis is proportionally enhanced by sequentially higher concentrations (10(-9) to 10(-4) molar) of benzyladenine. Ethidium bromide (1 microgram per milliliter) did not inhibit hydroxypyruvate reductase activity; thus, the enzyme synthesis does not appear to be controlled by organelle genes. Hydroxypyruvate reductase synthesis is inhibited by cycloheximide, cordycepin, and to a certain degree by actinomycin D. These data support the view of a close association between cytokinin action and gene expression.

Journal Article↗
PubMed1985

Localization of cytokinin biosynthetic sites in pea plants and carrot roots.

The biosynthesis of cytokinins was examined in pea (Pisum sativum L.) plant organs and carrot (Daucus carota L.) root tissues. When pea roots, stems, and leaves were grown separately for three weeks on a culture medium containing [8-(14)C]adenine without an exogenous supply of cytokinin and auxin, radioactive cytokinins were synthesized by each of these organs. Incubation of carrot root cambium and noncambium tissues for three days in a liquid culture medium containing [8-(14)C]adenine without cytokinin demonstrates that radioactive cytokinins were synthesized in the cambium but not in the noncambium tissue preparation. The radioactive cytokinins extracted from each of these tissues were analyzed by Sephadex LH-20 columns, reverse phase high pressure liquid chromatography, paper chromatography in various solvent systems, and paper electrophoresis. The main species of cytokinins detectable by these methods are N(6)-(Delta(2)-isopentyl_adenine-5'-monophosphate, 6-(4-hydroxy-3-methyl-2-butenyl-amino)-9-beta-ribofuranosylpurine-5'- monophosphate, N(6)-(Delta(2)-isopentenyl)adenosine, 6-(4-hydroxy-3-methyl-2-butenylamino)-9-beta-ribofuranosylpurine, N(6)-(Delta(2)-isopentenyl)adenine, and 6-(4-hydroxy-3-methyl-2-butenylamino)purine. On the basis of the amounts of cytokinin synthesized per gram fresh tissues, these results indicate that the root is the major site, but not the only site, of cytokinin biosynthesis. Furthermore, cambium and possibly all actively dividing tissues are responsible for the synthesis of this group of plant hormones.

Journal Article↗
PubMed1985

Separate resistances to arsenate and arsenite (antimonate) encoded by the arsenical resistance operon of R factor R773.

The arsenical resistance operon of R factor R773 was analyzed by subcloning and insertional inactivation. The operon was found to have two functional regions, the promoter-proximal region encoding resistance to arsenite and antimonate and the promoter-distal region encoding arsenate resistance. A unique 1.6-kilobase fragment was shown to be sufficient to encode arsenate resistance and produce arsenate extrusion from intact cells.

Arsenates↗
PubMed1984

Modification of cytokinins by cauliflower microsomal enzymes.

N(6)-(Delta(2)-isopentenyl)Adenine and N(6)-(Delta(2)-isopentenyl)adenosine were hydroxylated, respectively, to 6-(4-hydroxy-3-methyl-trans-2-butenylamino)purine and 6-(4-hydroxy-3-methyl-trans-2-butenylamino) -9-beta-ribofuranosylpurine in the presence of NADPH and the microsomal fraction from cauliflowers (Brassica oleracea L.). The hydroxylating reaction was completely inhibited by 10 millimolars metyrapone and partially inactivated by 10-minute treatment of the microsomal preparation with ethylene. The cytokinins were also dealkylated by the microsomal enzymes and formed adenine from cytokinin base and adenosine from cytokinin nucleoside. These results suggest that plant cytochrome P-450 is involved in the conversion of one type of cytokinin to another, and in the modification of cytokinin molecules.

Journal Article↗