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Biomedical subjects

M L Mace

Publications and source records attributed to M L Mace.

At least 37 records · Page 2Linked to original sources

Scanning electron microscopic examination of human asbestos bodies.

Asbestos bodies represent the product of the macrophage's attempt to detoxify inhaled asbestos fibers. The process of asbestos body maturation was examined by scanning electron microscopy of material isolated from lungs of former asbestos workers. The results suggest that the progression from a membrane-bound, smooth coating of the fiber, to the typically beaded form may be caused by cracking and subsequent erosion due to the inspiratory and expiratory forces of the lung on the asbestos body.

Asbestos↗

Variations in cell form and cytoskeleton in human breast carcinoma cells in vitro.

Cell form and cytoskeletal organization were investigated in 13 human breast carcinoma cell lines in vitro. Using tubulin antibodies and indirect immunofluorescence to detect the arrangement of cytoplasmic microtubules, three distinct cell phenotypes were recognized: (a) cells with extensive arrays of microtubules (type 1); (b) cells which were diffusely stained with microtubules apparent only near the cell margins (type II intermediate); and (c) cells in which individual microtubules could not be detected and only diffuse fluorescence was apparent (type II diffuse). Type I cells were flattened epithelial-like cells, much like normal mammary epithelial cells, which when stained with actin antibody displayed many brightly fluorescent parallel cables or "stress fibers." Many microtubules and microfilament bundles were observed in type I cells when examined by transmission electron microscopy. Type II cells were more rounded, often grew in multilayered colonies, and displayed fewer microtubules and microfilament bundles when examined by either immunofluorescence or electron microscopy. Type II cells ranged from very small rounded cells with diffuse tubulin and actin immunofluorescence (type II diffuse) to more flattened cells in which microtubules and actin cables were observed near the flattened cell margins (type II intermediate). Since all of the cells were derived initially from malignant metastatic lesions and some were tumorigenic when injected into athymic nude mice, we assume that they remained malignant in vitro. Thus, in human breast carcinoma cells in vitro, it is not possible to associate any specific cell morphology or cytoskeletal phenotype with cancer or metastasis in vivo. Whether or not these same conclusions hold for breast tumor cells in situ remains to be determined.

Actins↗

The chick ovomucoid gene contains at least six intervening sequences.

A 15-kilobase pair EcoRI chick DNA fragment, containing both the termination codon UGA and the 5'-portion of the structural ovomucoid gene, has been cloned in lambda phage Charon 4A by in vitro packaging. Restriction mapping and electron microscopic analyses of this cloned DNA have revealed that the structural ovomucoid gene sequences are separated by at least six intervening sequences.

Animals↗

Molecular structure and flanking nucleotide sequences of the natural chicken ovomucoid gene.

Five independent clones containing the natural chicken ovomucoid gene have been isolated from a chicken gene library. One of these clones, CL21, contains the complete ovomucoid gene and includes more than 3 kb of DNA sequences flanking both termini of the gene. Restriction endonuclease mapping, electron microscopy and direct DNA sequencing analyses of this clone have revealed that the ovomucoid gene is 5.6 kb long and codes for a messenger RNA of 821 nucleotides. The structural gene sequence coding Ifor the mature messenger RNA is split into at least eight segments by a minimum of seven intervening sequences of various sizes. The shortest structural gene segment is only 20 nucleotides long. All seven intervening sequences are located within the peptide coding region of the gene, and the sequences at the 5' and 3' untranslated regions of the mRNA are not interrupted by intervening sequences. The DNA sequences of the regions flanking the 5' and 3' termini of the gene have been determined. Thirty nucleotides before the start of the messenger RNA coding sequence is the heptanucleotide TATATAT, which is also present in a similar location relative to the chicken ovalbumin gene and other unique sequence eucaryotic genes. This sequence resembles that of the Pribnow box in procaryotic genes where a promoter function has been implicated. Seven nucleotides past the 3' end of the gene is the tetranucleotide TTGT, a sequence found to be present at identical locations as either TTTT or TTGT in other eucaryotic genes that have been sequenced. These conserved DNA sequences flanking eucaryotic genes may serve some regulator function in the expression of these genes.

Animals↗

The ovalbumin gene: cloning and molecular organization of the entire natural gene.

We report the analyses of recently cloned restriction fragments of the natural ovalbumin gene that overlap in part with previously cloned DNA fragments but extend further into the flanking sequences of the gene. These clones now permit us to identify the DNA sequence that codes for the 5' end of ovalbumin mRNA. Based on these and previous results, the molecular organization of the entire ovalbumin gene was established. The entire gene is composed of eight structural DNA sequences separated by seven intervening sequences that are not present in the mature mRNA. In addition, an ovalbumin gene clone has been obtained from a chicken gene library. Analysis of DNA isolated from this particular clone by molecular hybridization and electron microscopic mapping revealed that it contains the entire ovalbumin gene a single segment of DNA and its structure was consistent with that predicted from our physical map constructed from individually cloned fragments of the gene.

Animals↗

Assessment of attachment of Neisseria gonorrhoeae to HeLa cells by double radiolabeling.

Attachment of Neisseria gonorrhoeae to HeLa cells was assessed by a technique using double radioisotopic labeling. Piliated, virulent bacteria from colony type 2 attached to HeLa cells to a greater extent than nonpiliated, avirulent bacteria from colony type 4. Maximal attachment rates for bacteria from both colony types occurred during the early incubation periods at 37 degrees C, and the HeLa cells appeared saturated at 4 h. Attachment was maximum at pH 6.5 and dependent upon the multiplicity of infection. Treatment of the HeLa cells with trypsin diminished the degree of attachment, but this effect substantially disappeared by 24 h after trypsin treatment. Scanning electron microscopy revealed bacteria of colony types 2 and 4 adhered to the HeLa cell surface. Thin-section transmission electron microscopy showed that bacteria were associated with the surface of the HeLa cell but not ingested.

Cell Membrane↗

The natural ovalbumin gene contains seven intervening sequences.

EcoRI fragments of the natural ovalbumin gene were cloned and studied by hybridisation with mature ovalbumin mRNA, electron microscopy, restriction enzyme mapping and limited sequence analysis. The structural gene sequences coding for ovalbumin were found to be separated into eight sequentially orientated pieces by seven intervening sequences of various lengths.

Animals↗

Scanning-electron microscopy of chromosome aberrations.

This study is the first report of scanning-electron microscopy of isolated and purified metaphase chromosomes containing drug-induced aberrations. The technique reported allows high resolution topological examination of chromosomal aberrations which may pass undetected with conventional techniques.

Cell Line↗

Isolated metaphase chromosomes: scanning electron microscopic appearance of salt-extracted chromosomes.

Scanning electron microscopy of salt extracted, isolated metaphase chromosomes has been used to further the understanding of the role of proteins and DNA in chromosome architecture. Metaphase chromosome surfaces consist of numerous compact microconvules with an average diameter of 520 +/- 78 A which result from several orders of supercoiling. Treatment of isolated chromosomes with various salt concentrations ranging from 0.4 to 2.0 M NaCl revealed a longitudinal array of fibres within the chromatids as well as regions of coalesced nucleoprotein. The results demonstrate sensitivity of chromosome structure based on chromatin supercoiling, to progressive extraction of chromosomal proteins and DNA over the range of salt concentrations examined.

Cell Fractionation↗