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Biomedical subjects

T C Hsu

Publications and source records attributed to T C Hsu.

At least 199 records · Page 11Linked to original sources

Preparative-scale enzymic synthesis of D-[14C]ribulose 1,5-bisphosphate.

A procedure is described to prepare uniformly labelled D-[14C]ribulose 1,5-bisphosphate enzymically from uniformly labelled D-[14C]glucose through the coupled reactions catalysed by hexokinase (EC 2.7.1.1), glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44) and 5-phosphoribulokinase (EC 2.7.1.19). All reagents utilized in the method are commercially available. The procedure is a reliable preparative-scale method for synthesizing the dibarium salt of D-[14C]ribulose 1,5-biphosphate with a specific radioactivity up to 7 mCi/mmol and a purity near 90%. The final product was free of other 14C-labelled sugars, sugar phosphate esters, Pi and nucleotides.

Barium↗

Biochemical and cytological studies of bleomycin actions on chromatin and chromosomes.

Interaction of bleomycin with nuclei isolated from a variety of mammalian cells resulted in the release of nucleosomes. When isolated mononucleosomes (core plus linker) were re-treated with bleomycin, no further degradation of DNA occurred. The results suggest that the bleomycin cleavage sites in chromatin are present only in the linker region and that there are probably only one or two cleavage sites per linker. The repeat lengths of nucleosomal DNA released by bleomycin from nuclei of different species are different; this variability is considered to reflect the length of the linker. Incorporation of BrdU into DNA did not alter the bleomycin action on nucleosomes. When mitotic cells were held at metaphase for a prolonged period, bleomycin caused a gradual disintegration of chromosomes, although the bleomycin cleavage sites in metaphase chromosomes were found to be the same as those in interphase nuclei.

Animals↗

Studies in heterochromatin DNA: characterization of transcripts synthesized in situ from C-banded preparations.

The transcript of in situ RNA synthesis using Peromyscus eremicus cells, which had been treated with HCl followed by NaOH, proved to be rich in heterochromatin specifying sequences. Such transcript, referred to as heterochromatin RNA, can be characterized biochemically to probe the organization of heterochromatin DNA. It was found that the heterochromatin RNA of P. eremicus contained non-repetitive sequences covalently linked to repetitive sequences.

Animals↗

Are double minutes chromosomes?

Double minutes of a human breast cancer cell line revealed no centromeres by Cd banding. They cluster at the periphery of metaphase plates, usually encased in a matrix material. They move in anaphase passively with the chromosomes by attaching to the sides or ends of the chromosomes. The two "sister minutes" move to the same pole without separation. Such anomalous mitotic behavior suggests that double minutes are not chromosomes.

Anaphase↗

Cytogenetic toxicity of gentian violet and crystal violet on mammalian cells in vitro.

The cytogenetic toxicity of gentian violet in Chinese hamster CHO cells in vitro has been studied by analyzing (1) squash preparations from direct fixation for recording mitotic anomalies and (2) air-dried preparations (with colcemid--hypotonic pretreatments) for recording metaphase chromosome aberrations. It was concluded that this compound is a mitotic poison as well as a clastogen in vitro. Its clastogenic property was confirmed in 5 other different mammalian cel types. 10 samples of different gentian violet and crystal violet were surveyed and all were shown to be clastogenic. Unless in vivo studies prove otherwise, gentian violet and crystal violet should be regarded as biohazardous substances.

Animals↗

Induced Robertsonian fusions and tandem translocations in mammalian cell cultures.

Cultures of a cattle cell line and a Peromyscus eremicus cell line recovering from a pulse-treatment with mitomycin C, actinomycin D, 33258 Hoechst, and nitrosoguanidine exhibited translocations between chromosomes at the centromeric regions (Robertsonian fusions) as well as between centromere and telomere and between telomeres (tandem translocations). The frequency of Robertsonian fusions was found to be dose-dependent and duration-dependent with the mitomycin treatment. Biarmed chromosomes resulting from fusions may be monocentric or dicentric. Analyses of clones isolated from treated cells suggested that fused chromosomes may perpetuate in the cell populations.

Animals↗

Combination of silver and fluorescent staining for metaphase chromosomes.

A convenient and reliable method for simulatneous visualization of silver staining (Ag-NOR) of the nucleolus organizers and fluorescent bandings in metaphase chromosomes is described. Studies employing this combined procedure on human chromosomes revealed that the Ag-NOR patterns may be characteristic for each chromosome of each individual.

Chromosomes↗

Visualization of centriole-bodies using silver stain.

Duplicate staining of human metaphase chromosomes, first with Giemsa followed by silver, revealed the presence of two small silver-stained bodies not seen in the Giemsa stained metaphases. Similar bodies were subsequently found in the metaphases of several animal groups. The size, structure, spatial relationships to the nucleus, behavior throughout the cell cycle, and apparent universal presence of these bodies suggest that they are either centrioles or associated centriolar structures. These centriole-bodies can be seen throughout the cell cycle, even in hypotonically spread C-metaphase chromosome plates. The silver stain procedure allows enough resolution to distinguish parent and daughter centriole-bodies at interphase, thus permitting visualization of the replication, maturation and separation stages of these bodies with the light microscope.

Animals↗

Cytogenetic assays of chemical clastogens using mammalian cells in culture.

A protocol is described for cytogenetic assays of chemical mutagens using mammalian cells in vitro. The system employs continuous drug treatment (3 concentrations) for up to 8 h and recovery-cell populations after pulse treatments with a high dose. Both direct fixation (for recording spindle anomalies in anaphase) and colcemid-hypotonic fixation (for reading metaphase chromosome aberrations) are used in order to estimate the effects of an agent as a mitotic poison and as a clastogen respectively. Some DNA intercalating dyes (acridine orange, quinacrine mustard, neutral red) were found to be highly clastogenic whereas others (quinacrine dihydrochloride, 33258 Hoechst) are not.

Cell Line↗

Identification of nucleolus organizer regions (NORs) in normal and neoplastic human cells by the silver-staining technique.

Silver nitrate has been used to demonstrate the chromosomal location of ribosomal cistrons in nine tissue-culture lines derived from human tumors of various pathological origins. Control individuals have a particular modal number (range 7--10) of D- and G-group chromosomes stained with silver. In the controls, 96.2% of the D- and G-group chromosomes that have a stalk show silver staining, while no relationship can be seen in acrocentric chromosomes without stalks. The tumor cells, whose modal chromosome numbers range from 42 to 68, possess variable numbers of acrocentrics (11--18). The number of chromosomes stained with silver, however, remained at control levels (range, 6--9). These data indicate that, in humans, silver staining may not identify all NORs that contain structural ribosomal genes.

Cell Line↗

Chromosomes of Peromyscus (rodentia, cricetidae). VI. The genomic size.

In the genus Peromyscus cells of all species contain 48 chromosomes; however, the fundamental number varies from 56 (P. Crinitus, P. boylei) to 96 (P. eremicus). In some cases biarmed chromosomes are the result of pericentric inversions, while in others they are the result of addition of large amounts of constitutive heterochromatin. Flow microfluorometric DNA-per-cell determinations demonstrated that in some species (P. eremicus) the genome is increased by 36% over the amount of DNA found in most mammalian species. Studies of unique karyotypes with increased amounts of DNA added as constitutive heterochromatin may ultimately help in the elucidation of the mechanisms involved in karyotype evolution and speciation.

Animals↗

Renal bisection.

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Adolescent↗

Adriamycin-induced chromosome damage: elevated frequencies of isochromatid aberrations in G2 and S phases.

Analysis of chromosomes from cells treated with adriamycin during G2 and S phases showed a high frequency of isochromatid-type of breaks, in addition to the expected chromatid breaks. These are interpreted as independent breaks on sister chromatids because of preferential effects of the drug in specific chromosome regions. The break points are likely to be different, but morphologically such breaks would be indistinguishable from isochromatid or chromosome breaks.

Cell Line↗