A characterization study of a cell line (BS/BEK) from bovine embryo kidney.

A cell line (BS/BEK) which was obtained from bovine embryo kidney tissue, when studied at its 140th passage level it showed the following properties: 1. An epithelial-like morphology, possessing a heteroploid karyotype with a modal chromosome number ranging between 70 and 75 chromosomes. 2. It failed to produce tumors in mice and in hamster. 3. It was shown to be ready susceptible to the replication of several viral agents originated from a variety of animal species. 4. It was not contaminated by mycoplasma or other bacterial spp.

Study of fibronectin expression in tumour cells by dot-blot and in situ hybridization: quantitative evaluation by image analysis.

An Image Analysis program was used for the quantitative evaluation and comparison of the fibronectin (FN) mRNA detected by dot-blot and in situ hybridization in different cell lines. These techniques were applied for the evaluation of FN mRNA synthesized by human normal fibroblasts (Flow 7000) and by four tumour-derived cell lines (HeLa, epithelioid carcinoma; 8387, fibrosarcoma; RD, rhabdomyosarcoma; SK Hep-1, hepatocarcinoma). Dot-blot analysis showed that the cell types analysed synthesize different levels of FN mRNA. Flow 7000 are the highest producers while HeLa the lowest. In situ hybridization confirmed these results and furthermore showed that while Flow 7000, 8387 and HeLa cells synthesized homogeneous levels of FN mRNA, RD and SK Hep-1 could be subdivided into two populations expressing high or low levels of FN mRNA. The combined analysis of dot-blot, in situ hybridization and Image Analysis allowed the quantitation of the number of FN mRNA molecules expressed by single cells. This approach is therefore an invaluable tool when evaluating mRNA expression in heterogeneous cell populations like tumour-derived cell lines, during cell cycle or in histological tissue sections.

General characteristics and viral susceptibility of a newborn pig kidney (NPK) continuous culture.

We have developed a fibroblastic-like continuous culture of newborn pig kidney (NPK). The current cell line was serially passaged 160 times and appeared to be well suited for production and assay of a number of viruses affecting pigs, such as pig parvovirus, pseudorabies and transmissible gastroenteritis. The cell line appeared aneuploid, with a modal chromosome number of 36 and induced tumors, classified as fibrosarcoma, in athymic mice.

Malformation syndrome with t(2;22) in a cancer family with chromosome instability.

A de novo unbalanced t(2;22)(q37;q11.2) [corrected], resulting in the deletion of the 22pter-q11 and 2q37-qter regions, was observed in a 12-year-old girl born with a congenital malformation syndrome and later displaying signs of neurologic impairment. Some of the clinical signs observed appear to overlap those found in subjects monosomic in the 22q11 region affected by the DiGeorge syndrome. The chromosomal rearrangement observed may be related to a familial cytogenetic instability that also gives rise to sustained cancer predisposition.

Cytogenetic instability in a family with gastric cancer recurrence.

An index case with a congenital malformation syndrome enabled detection of a family that had a previous history of spontaneous abortuses and recurrence of neoplasia through three generations. Cytogenetic analysis performed on lymphocytes from 11 subjects in the second and third generation showed karyotypic alterations in both tumor bearers and apparently normal subjects. Chromosome variations consisted of: spontaneous chromosome fragility; chromosome translocations; polymorphisms in the heterochromatic regions in chromosomes Y, #1, #16, #22. The inheritance pattern of all chromosome rearrangements and heteromorphisms observed was established starting with the second generation, and the contribution of specific individuals was identified. Although the relationship between chromosomal instability and predisposition to gastric cancer does not appear to be coincidental, no specific chromosome alteration in normal somatic cells was shared by all members of the family who developed or are at risk of developing tumors.

Chromosomal location by in situ hybridization of the human Sau3A family of DNA repeats.

The Sau3A family is a human, clustered, highly repetitive, GC-rich DNA family. In situ hybridization studies with a plasmid carrying a Sau3A monomer as a probe have shown that Sau3A sequences are preferentially concentrated in the heterochromatic regions of human acrocentric chromosomes (D and G groups, both in pericentromeric regions and in cytological satellites) and in pericentromeric heterochromatin of chromosome 1. The same chromosomal locations were observed by using as probes two recombinant phages which carry Sau3A-positive genomic sectors. The two sectors differ for the relative proportions of monomer and multiples of Sau3A repeats, which show different extents of homology to the cloned monomer, and for the presence, in one of the two, of a small amount of an unrelated repeat (alphoid DNA). The similarity of the results obtained with the three probes suggests that heterogeneous Sau3A repeats share the same chromosomal localizations and that the two analyzed genomic sectors may not contain significant amounts of repetitive DNAs other than the Sau3A family. A comparison between the chromosomal locations of Sau3A and EcoRI families of repeats has confirmed that each family is characterized by specific chromosomal locations and that single heterochromatic regions may contain both.

The study of X-rays and TCDD effects on satellite associations may suggest a simple model for application in environmental mutagenesis.

Satellite associations were used as parameters to test nucleolar organizer activity. Assuming that toxic and/or mutagenic agents may affect the ribosomal genes, satellite associations in human lymphocytes were analysed following exposure to X-rays and compared with the satellite association pattern of subjects exposed to TCDD. A significant decrease in the satellite association frequency in D group chromosomes was found both in irradiated lymphocytes and in subjects exposed to Dioxin. The findings seem to be in accordance with the hypothesis based on random damage of functional nucleolar organizing regions.

Satellite associations and silver staining in a case of multiple G and D variants.

Satellite associations and silver staining were analyzed in a normal woman carrying three "s" variants, on chromosomes 13, 21, and 22. Six of the acrocentric chromosomes were identified and a positive correlation between the parameters of satellite association frequency and positive silver staining was found for each chromosome. These parameters seem to depend on the presence and size of a secondary constriction and are unaffected by stallelite size.

Cytotoxic effects of phallolysin on a human heteroploid cell line.

The effect of Phallolysin on cellular growth, macromolecular biosyntheses and cellular membrane structure was analysed using cells from the EUE line. Concentrations of the toxin that do not affect cellular growth, as determined by plating efficiency, have no effect on RNA or protein synthesis, but stimulate DNA synthesis. The doses of Phallolysin that inhibit cell survival do not affect macromolecular biosyntheses, but greatly increase the percentage of cells stainable with Trypan blue after 1 hour of incubation. At the same dose of the toxin the cells, analysed by electron microscopy, show increased vacuolization indicating an alteration of the membrane apparatus.

Cytotoxic effects of peptichemio on synchronized EUE cells.

The effect of Peptichemio (PTC) on cellular growth and on macromolecular syntheses was analyzed through the cell cycle of EUE cells. The cell response to the various treatments was measured by determining plating efficiency, growth rates and incorporation of labelled precursors of DNA, RNA and protein syntheses. Three maximum inhibition points were found on cell survival, one corresponding to the early G1, another to the middle S and a last one to the late G2. Parallel experiments of incorporation of labelled precursors of DNA, RNA and proteins revealed an effect only on DNA during the early and middle S phase.

The organization of DNA sequences in the mouse genome.

Analysis of the organization of nucleotide sequences in mouse genome is carried out on total DNA at different fragment size, reannealed to intermediate value of Cot, by Ag+--Cs2SO4 density gradient centrifugation.--According to nuclease S-1 resistance and kinetic renaturation curves mouse genome appears to be made up of non-repetitive DNA (76% of total DNA), middle repetitive DNA (average repetition frequency 2X10(4) copies, 15% of total DNA), highly repetitive DNA (8% of total DNA) and fold-back DNA (renatured density 1.701 g/ml, 1% of total DNA).--Non-repetitive sequences are intercalated with short middle repetitive sequences. One third of non-repetitive sequences is longer than 4500 nucleotides, another third is long between 1800 and 4500 nucleotides, and the remainder is shorter than 1800 nucleotides.--Middle repetitive sequences are transcribed in vivo. The majority of the transcribed repeated sequences appears to be not linked to the bulk of non-repeated sequences at a DNA size of 1800 nucleotides.--The organization of mouse genome analyzed by Ag+--Cs2SO4 density gradient of reannealed DNA appears to be substantially different than that previously observed in human genome using the same technique.