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Biomedical subjects

R J Wang

Publications and source records attributed to R J Wang.

At least 37 records · Page 2Linked to original sources
PubMed1987

The autoimmune antigen Me is distinct and related to undifferentiated connective tissue disease.

Using prototype Me serum, a new autoantibody-antigen system has been identified by Ouchterlony immunodiffusion and indirect immunofluorescence. Although immunologically distinct, the Me antigen has physiochemical and biochemical properties similar to those of the Sm antigen. Immunoblot assays indicate that Me sera commonly recognize 4 peptides of molecular weights approximating 100K, 65K, 21K, and 16K. The last of these may be identical to the D peptide recognized by Sm antibodies. The Me antigen may be associated with the RNP-Sm macromolecular complex. Me-positive patients have few clinical symptoms, and the most common diagnosis is undifferentiated connective tissue disease.

Antigen-Antibody Complex↗
PubMed1984

Lysine tRNA and cell division: a G1 cell cycle mutant is temperature sensitive for the modification of tRNA5Lys to tRNA4Lys.

Ts-694 is a temperature sensitive mutant of hamster cells which is blocked in the G1 phase of the cell cycle at the restrictive temperature of 39 degrees. A comparison of the Lys-tRNA isoacceptors by RPC-5 chromatography showed a decrease in tRNA5Lys and an increase in tRNA4Lys at 39 degrees. This was identical to the changes seen in confluent cultures at the permissive temperature of 33 degrees. These Lys-tRNA changes were not seen in ts-694 cells blocked in G1 by isoleucine deficiency, nor in two other G1 ts mutants at the restrictive temperature. Cells trapped in S phase by a thymidine block also contained decreased levels of tRNA4Lys when raised to 39 degrees. Both tRNA4Lys levels and cell division increased when the cells were returned to the permissive temperature. An in vitro assay was established for the modification of tRNA5Lys to tRNA4Lys with tRNA6Lys and tRNA2Lys as intermediates. The first reaction is the synthesis of tRNA6Lys which involves the introduction of a modified uridine at the third position of the anticodon. Extracts of 694 cells grown at 33 degrees were able to modify rat liver [3H] tRNA5Lys to tRNA6Lys and tRNA4Lys in vitro when assayed at 25 degrees but not at 39 degrees. Extracts of Balb/c 3T3 cells, however, were more active at 39 degrees than at 25 degrees showing that the normal enzyme is not temperature sensitive. Ts-694 cell tRNA, isolated from cells grown at 33 degrees was aminoacylated at both 25 degrees and 39 degrees with rat liver synthetases. tRNA4Lys was present at both temperatures indicating that ts-694 cells do not contain a temperature sensitive tRNA4Lys.

Animals↗
PubMed1984

Cell division cycle in mammalian cells. VIII. Mapping of G1 into six segments using temperature-sensitive cell cycle mutants.

The G1 blocks in three temperature-sensitive (ts) Syrian hamster cell-cycle mutants have been mapped in relation to other G1 landmarks. Two mutants reported here, ts-559 and ts-694, show defective progression only in G1. When shifted from the permissive temperature of 33 degrees C to the non-permissive temperature of 39 degrees C, G1 cells of these two mutants show no further cell cycle progression, while cells in S, G2 and mitosis progress through the cell cycle but become blocked after entering G1. The two mutants complement each other, and also complement the previously reported mutant ts-550C with blocks in both G1 and G2 of the cell cycle. The locations of the G1 blocks in both ts-559 and ts-694 are before the hydroxyurea arrest point. The G1 ts point in ts-694 is prior to the isoleucine deprivation and serum starvation points, while the G1 block in ts-559 is after the serum starvation point but before the isoleucine block. Other G1 block points which have been reported are in mutants of different species and isolated in different laboratories, causing difficulties for relative positioning of the blocks in G1. The mutants for mapping in this study have been isolated from the same cell line. The G1 ts arrest points of ts-559 and ts-694, and that found in ts-550C, together with nutritional deprivations and metabolic inhibitors, provide seven reference points which divide G1 into six segments, each of which is bracketed by two adjacent points: mitosis, ts-694 block, serum starvation arrest point, ts-559 block, isoleucine deprivation arrest point, ts-550C block, hydroxyurea or excess-thymidine arrest segment.

Animals↗
PubMed1983

Studies on cell division in mammalian cells. VII. A temperature-sensitive cell line abnormal in centriole separation and chromosome movement.

A temperature-sensitive Syrian hamster mutant cell line, ts-745, exhibiting novel mitotic events has been isolated. The cells show normal growth and mitosis at 33 degrees C, the permissive temperature. At the nonpermissive temperature of 39 degrees C, mitotic progression becomes aberrant. Metaphase cells and those cells still able to form a metaphase configuration continue through and complete normal cell division. However, cells exposed to 39 degrees C for longer than 15 min can not form a normal metaphase spindle. Instead, the chromosomes are distributed in a spherical shell, with microtubules (MT) radiating to the chromosomes from four closely associated centrioles near the center of the cell. The cells progress from the spherical monopolar state to other monopolar orientations conical in appearance with four centrioles in the apex region. Organized chromosome movement is present, from the spherical shell state to the asymmetrical orientations. Chromosomes remain in the metaphase configuration without chromatid separation. Prometaphase chromosome congression appears normal, as the chromosomes and MT form a stable monopolar spindle, but bipolar spindle formation is apparently blocked in a premetaphase state. When returned from 39 degrees to 33 degrees C, the defective phenotype is readily reversible. At 39 degrees C, the mitotic abnormality lasts 3-5 h, followed by reformation of a single nucleus and cell flattening in an interphase-like state. Subsequent cell cycle events appear to occur, as the cells duplicate chromosomes and initiate a second round of abnormal mitosis. Cell cycle traversion continues for at least 5 d in some cells despite abnormal mitosis resulting in cells accumulating several hundred chromosomes.

Animals↗
PubMed1982

Studies on cell division in mammalian cells: VI. A temperature-sensitive mutant blocked in both G1 and G2 phases of the cell cycle.

A temperature-sensitive mammalian cell cycle mutant with blocks in G1 and G2 phases of the cell cycle has been isolated in culture. When shifted from the permissive temperature of 33 degrees C to the nonpermissive temperature of 39 degrees C, the fraction of cells initiating DNA synthesis as well as the fraction of cells entering mitosis decreased rapidly. Combined cytophotometric and autoradiographic analysis on the cells at 39 degrees C showed that G1 cells, with the exception of those in late G1, were arrested in that phase. Cells is S phase at the time of temperature shift, together with the late g1 cells which subsequently entered S, continued through S into G2, but were blocked in that phase of the cell cycle and unable to initiate mitosis. Those cells already in mitosis completed cell division at 39 degrees C. The G1 block point of ts-550C was found to be located after the serum starvation and isoleucine deprivation arrest points, approximately 3 h before initiation of DNA synthesis.

Aged↗
PubMed1981

Tenotomy of the avian anterior latissimus dorsi muscle. I. Effect of age on fiber-type transformation and regeneration from the stump in chicks.

The chick's anterior latissimus dorsi muscle (ALD) was tenotomized at its origin at either 1 day or 4 weeks of age, and investigated histochemically and ultrastructurally at intervals thereafter to determine whether muscle fiber-type transformation from a slow to a twitch type is greater in young birds than older birds. No transformation of fiber type occurred in either procedure, but a new muscular connection regenerated between the scar tissue at the end of the original tenotomized stump and the former origin. This regenerated muscle had a mosaic pattern of fiber types, as demonstrated by myofibrillar ATPase activity, and contained predominantly fast fibers, as contrasted with controls or the tenotomized portion, which contained predominantly slow tonic muscle fibers. The regenerated portion contained muscle spindles. The original portion of the tenotomized muscle was indistinguishable from the control muscle. These responses of the chick ALD to tenotomy are quite different from those in the pigeon, which are reported in the following study.

Adenosine Triphosphatases↗
PubMed1981

A differential staining technique for simultaneous visualization of mitotic spindle and chromosomes in mammalian cells.

A new technique has been devised for staining the mitotic spindle in mammalian cells while preserving spindle structure and chromosome number. The cells are trypsinized and fixed with a 3:1 methanol:acetic acid solution containing 4 mM MgCl2 and 1.5 mM CaCl2 at room temperature. The cells are then placed on slides and treated with 5% perchloric acid before staining with a 10% acetic acid solution containing safranin O and brilliant blue R. The preserved spindles appear dark blue against a light cytoplasmic background with chromosomes stained bright red. Individual chromosomes and chromatids are clearly visible. Positioning of the chromosomes relative to the spindle apparatus is readily ascertained allowing easy study of mitotic spindle and chromosome behavior.

Animals↗
PubMed1978

Identification of hydrogen peroxide as a photoproduct toxic to human cells in tissue-culture medium irradiated with "daylight" fluorescent light.

Hydrogen peroxide, lethal for human cells, is produced in Dulbecco's modified Eagle's tissue culture medium when exposed to "daylight" fluorescent light. Addition of pure H2O2 and use of the enzyme catalase demonstrate that about 40% of the toxicity in irradiated medium is due to generated peroxide. Riboflavin and tryptophan, or riboflavin and tyrosine, are the components necessary for formation of lethal levels of H2O2 during light exposure.

Catalase↗
PubMed1976

Effect of room fluorescent light on the deterioration of tissue culture medium.

A major cause of tissue culture medium deterioration is exposure to room fluorescent light. Riboflavin and tryptophan present in Dulbecco's modified Eagle's minimum essential medium, when exposed to light, yield toxic photoproducts responsible for loss of the ability of the medium to support clonal growth of human, mouse and Chinese hamster cell lines. Procedures for minimizing medium deterioration are discussed.

Cell Line↗
PubMed1976

A novel temperature-sensitive mammalian cell line exhibiting defective prophase progression.

A temperature-sensitive mammalian cell line has been isolated which grows and divides normally at the permissive temperature of 33 degrees C. When incubated at 39 degrees C, the nonpermissive temperature, interphase cells continue to enter a prophase-like state. Chromatin-like material condenses and coalesces into dark-staining clumps rather than into discernible chromosomes. Disappearance of the nuclear boundary is observed, but re-formation of the boundary around the clumps fails to occur. In corporation of labeled precursors reveals a decrease in protein synthesis which is accompanied by a slower decrease in DNA synthesis. Approximately 0.2% of the mutant cells revert in their capability of growth and cell division at 39 degrees C. These "revertants" are found to contain a higher number of chromosomes. The isolation of this mutant is based on the initial observation that the cells become rounded at the nonpermissive temperature. The cell-rounding process characteristic of mitotic cells should serve as a useful marker in the isolation of mitotic mutants.

Animals↗