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Biomedical subjects

O J Miller

Publications and source records attributed to O J Miller.

At least 109 records · Page 6Linked to original sources

Serologic detection of a y-linked gene in xx males and xx true hermaphrodites.

To test the hypothesis that H-Y antigen (present on both somatic and germ cells in normal males but not normal females) is essential for testicular differentiation, we studied four XX males and three XX true hermaphrodites. Blood cells from six subjects and cultured gonadal fibroblasts from a seventh expressed H-Y antigen. Since expression of this antigen requires the presence of a gene normally carried by the Y chromosome, this gene, and perhaps additional Y chromosomal material, should have been present in the genome of these subjects. In one patient this presence is accounted for by a Y-to-X translocation, detectable by chromosome banding. In another a normal Y chromosome was present in a minor population of cells. In the remaining five, no karyotypic abnormality was detectable. Immunologic detection of H-Y antigen is a sensitive test for the presence of the Y chromosome or of its male-determining segment.

Adult↗

Detection of nucleolus organizer regions in chromosomes of human, chimpanzee, gorilla, orangutan and gibbon.

Nucleolus organizer regions were detected by the Ag-AS silver method in fixed metaphase chromosomes from human and primates. In the human, silver was deposited in the secondary constriction of a maximum of five pairs of acrocentric chromosomes: 13, 14, 15, 21 and 22. The chimpanzee also had five pairs of acrocentric chromosomes stained, corresponding to human numbers 13, 14, 18, 21 and 22. A gibbon had a single pair of chromosomes with a secondary constriction, which corresponded to the nucleolus organizer region. In each case the Ag-AS method detected the sites which have been shown by in situ hybridization to contain the ribosomal RNA genes. An orangutan had eight pairs of acrocentric chromosomes stained with Ag-AS, probably corresponding to human numbers 13, 14, 15, 18, 21 and 22, plus two others. Two gorillas had silver stain over two pairs of small acrocentric chromosomes and at the telomere of one chromosome 1. The larger gorilla acrocentric chromosomes had no silver stain although they all had secondary constrictions and entered into satellite associations.

Animals↗

Antibodies to histones and histone-histone complexes: immunochemical evidence for secondary structure in histone 1.

Highly specific antibodies were raised to histone 1 (H 1) and the histone complexes H32-H42 AND H2A-H2B, isolated by salt extraction. Antibody to H1 could detect irreversible conformational changes in acid- or urea-treated H1. The antibodies showed different reactivities with chromosomes as compared to antibodies in acid-extracted histones and should be useful in studies of native chromatin and chromosome structure.

Antibodies↗

5-methylcytosine in heterochromatic regions of chromosomes in Bovidae.

The centromeric regions of cattle, goat and sheep chromosomes bind anti-5-MeC as revealed by immunofluorescence technique, indicating concentration of 5-MeC at these heterochromatic regions. The centromere of the submetacentric X of cattle remains nearly unstained and so do the centromeres of the acrocentric X chromosomes in goat and sheep. The short arm of the cattle Y exhibits strong anti-5-MeC binding whereas the tiny Y chromosomes of goat and sheep contain no brightly fluorescent material.

Animals↗

The role of trypsin in the pre-treatment of chromosomes for Giemsa banding.

The role of trypsin in the elicitation of G-banding on human chromosomes was studied in two separate laboratories. Enzyme activity and ability of trypsin to chelate calcium were manipulated by dilution of the treatment solution, and by inhibition with diisopropylphosphofluoridate, diphenylcarbamyl chloride, or soybean trypsin inhibitor. In all cases, chromosomes were affected in proportion to the enzyme activity of the treatment solution rather than the ability of the solution to bind calcium. It is concluded that calcium chelation is not sufficient to explain G-banding trypsin, but that proteolytic activity is required.

Azure Stains↗

Localization of 5-methylcytosine in human metaphase chromosomes by immunoelectron microscopy.

Human metaphase chromosomes, fixed in methanol-acetic acid, were ultraviolet irradiated to produce single-stranded regions of chromosomal DNA and treated with anti-5-methylcytidine. Using an immunoperoxidase procedure, regions of antibody binding were readily visualized by light microscopy in the centromeric heterochromatin regions of chromosomes 1, 9, 16, the short arm of chromosome 15, and in the distal portion of the Y. Electron microscopic visualization of the same whole mount chromosome preparations transferred to formvarcoated grids revealed additional details of the distribution and arrangement of 5-methylcytosine. A helical arrangement of 5-methylcytosine residues was seen below the centromere of chromosome 1. The Y chromosome showed a concentration of 5-methylcytosine residues on the distal long arm, and in areas just below and slightly above the centromere. In all the above chromosomes, especially chromosome 15, additional 5-methylcytosine residues were detected as isolated foci along the arms. Our findings support the concept that clusters of similar purine or pyrimidine residues exist along the arms of condensed metaphase chromosomes, with the possibility that concentrations of 5-methylcytosine residues might have been enhanced at the surface of the chromosomes during the condensation process.

Antibodies↗

Expression of human and suppression of mouse nucleolus organizer activity in mouse-human somatic cell hybrids.

Most mouse-human somatic cell hybrids show preferential loss of human chromosomes, absence of human 28S ribosomal RNA, and suppression of human nucleolus organizer activity, as visualized by the Ag-AS silver histochemical stain. In contrast, the mouse-human hybrids studied here show preferential loss of mouse chromosomes. The hybrids were made by fusion of HT-1080-6TG human fibrosarcoma cells with BALB/c mouse peritoneal macrophages or strain 129 mouse teratocarcinoma cells. The Ag-AS staining method shows nucleolus organizer activity of chromosomes 13, 14, 15, 21 (rarely), and 22 in the human parent and chromosomes 12, 15, 16 (rarely), and 18 in the BALB/c mouse parent. In the hybrid cells the human nucleolus organizer regions are active, as shown by Ag-AS staining and involvement in "satellite association." The mouse nucleolus organizer regions are not stained by the Ag-AS method even though mouse chromosomes 12, 15, and 18 are present in the BALB/c hybrids and at least one copy of each mouse chromosome is present in the teratocarcinoma-derived hybrids. Thus, in these mouse-human hybrids, unlike those that lose human chromosomes, only human nucleolus organizer activity is expressed, and mouse nucleolus organizer activity is suppressed.

Animals↗

Erythroid cell differentiation: murine erythroleukemia cell variant with unique pattern of induction by polar compounds.

The murine-virus-infected erythroleukemia cell system provides an opportunity to examine regulatory mechanisms controlling cytodifferentiation. A cloned cell line (DR10c3) resistant to the erythropoiesis-inducing effect of dimethylsulfoxide (Me2SO) was isolated from the Me2SO-sensitive line DS19. DR10c3 is characterized as follows: (1) the uptake of [3H]Me2SO is similar to that in DS19; (2) cell growth with and without Me2SO is similar to that of DS19; (3) resistance is relatively stable; (4) the karyotype of DR10c3 reveals an average loss of five chromosomes per cell, but is otherwise similar to that of DS19; (5) total protein and globin synthesis by cells cultured 4 days with or without Me2SO is similar to these syntheses in DS19 cultured without Me2SO; (6) virtually no globin mRNA is detectable after 3 days in Me2SO, as assayed both by RNA-complementary DNA hybridization and by the heterologous cell-free protein-synthesizing system; (7) other polar compounds, N-methylpyrrolidinone, 1-methyl-2-piperidone, N, N-dimethylacetamide, and N-methylacetamide, induce erythroid differentiation in DR10c3, and the accumulation of alpha- and beta-globin chains is indistinguishable from that in DS19; and (8) the concentration optima for induction of differentiation by all these compounds are identical for DR10c3 and DS19.

Acetamides↗

Q- and C-band chromosome markers in inbred strains of Mus musculus.

Differences in the number of chromosomes with secondary constrictions and in the size of the C-band region on certain chromosomes have been observed among the following inbred strains of Mus musculus: C57BL/10J, C57BR/cdJ, DBA/1J, CBA/J, BALB/cJ, and AKR. These differences are useful as indicators of the location of rRNA genes and as normal chromosome markers. The size of each C-band region appears to remain constant over many generations. Only one probable change in the size of a C-band region was found.

Animals↗

Further evidence of X-linkage of hypoxanthine phosphoribosyl-transferase in the mouse.

A variant of the HPRT-A9 mouse cell line was fused with wild type diploid mouse bone marrow cells to obtain an HPRT+ line. The unique chromosomal features of the A9 parent, including the presence of a t(X;3) translocation and the absence of normal chromosomes 15,16, 17, 18, and the X, have permitted use of this intraspecific hybrid for chromosome mapping. Back-selection of hybrid cells in 8-azaguanine for loss of HPRT resulted in the loss of the Xchromosome derived from the diploid parent, providing evidence of the X-linkage of the HPRT locus.

Animals↗