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Biomedical subjects

K L Sternes

Publications and source records attributed to K L Sternes.

6 recordsLinked to original sources

Use of a Salmonella typhimurium hilA fusion strain to assess effects of environmental fresh water sources on virulence gene expression.

Many fruits and vegetables are irrigated with water from rivers, lakes and even wastewater systems. Irrigation may be a route for the introduction of Salmonella. Our objectives in this study were to determine survivability and virulence expression in a strain of Salmonella typhimurium when exposed to environmental water sources. Virulence expression was measured using a beta-galactosidase assay on a hilA:lacZY fusion strain of S. typhimurium. Water samples for environmental impact studies were taken from a local pond and specific sites along the Rio Grande River, which serves as a source of irrigation water in southern Texas. There was a significant difference (p<0.05) of virulence expression among the water sites. Certain regions along the Rio Grande River yielded greater amounts of beta-galactosidase activity than others. All sites yielded at least a two-fold greater virulence response than S. typhimurium grown in brain heart infusion. Salmonella survivors were enumerated as colony forming units (CFU)/ml as plated on a selective medium for the duration of 1 week and beta-galactosidase assays were performed to determine a possible correlation between culturable cells and virulence gene expression. Bacterial cells remained viable but decreased after 7 days incubation. In conclusion, water sampled at specific locations and at different times water samples exhibited differences in virulence expression in S. typhimurium.

Agriculture↗

Satellite I DNA in transformed rat cells.

P93-50, a 93-basepair (bp) repetitive DNA sequence from rats, was hybridized to transformed sublines of rat endothelial origin. The sequence hybridized at or near the centromeres of most but not all chromosomes in two transformed cell lines and three single-cell derived cultures. The hybridization signal was also frequently present at the telomeres. These cell lines have a highly aberrant karyotype including dicentric and multicentric chromosomes; however, even though this sequence labeled the centromere regions of some chromosomes, it did not hybridize with the telomere regions of the cell line XC, which rarely shows any dicentrics. Apparently, the telomere signals represent prematurely separating, inactive, terminal centromeres. The p93-50 sequence does not influence the timing of centromere separation, nor it is necessary for formation of heterochromatin.

Animals↗

Cytogenetic variability and kinetochore proteins. Comparison among populations derived from single-cell cultures.

This study reports comparative changes in five sub-lines obtained from a transformed culture of rat cerebral origin. Two of the lines were obtained from the original cell population while three others were raised from single-cell cultures. The comparative study was carried out on the DNA content and several cytogenetic parameters including variability in chromosome number, anaphase bridges, acentric fragments, chromosomes without detectable kinetochore proteins, and the frequency of micronuclei in these five lines. All cell lines, including the single-cell-derived clones expressed considerable variability in all aspects. One interesting aspect is the evolution of a chromosome with compound centromere, which is present only in two cell lines. The data indicate that the clone derived from a single cell does not maintain uniformity and even single cells have some sort of inherent potential to generate extreme variability. Some numerical variability may be attributed to a new phenomenon of a lack of kinetochore proteins seen on some chromosomes.

Anaphase↗

Centromeres without kinetochore proteins. Another mechanism for origin of aneuploidy in neoplasia.

Centromeres of all chromosomes in normal cells exhibit kinetochore proteins detectable by antikinetochore antibodies. The present communication reports that some chromosomes in a transformed cell line of rat cerebral origin fail to deposit kinetochore proteins at their centromeres. These chromosomes may not undergo normal anaphase segregation and may be either lost or enter one or the other daughter cell. The observation that some chromosomes may be without detectable kinetochore proteins suggests a noval mechanism for origin of aneuploidy in transformed and neoplastic cells.

Aneuploidy↗

Centromere structure and function in neoplasia.

The mammalian centromere plays an essential role in maintenance of diploidy in the cell. It is therefore imperative that we understand the structure and function of the mammalian centromere in order to plan strategy to control the incidence of aneuploidy and resultant malformations of the nonneoplastic as well as neoplastic tissues. Even though considerable information is available about the structure and some functional aspects of centromeres of lower eukaryotes such as yeast, the structure of the mammalian centromere is still a matter of conjecture limited to an understanding of the base composition of the alphoid sequences putatively located in the centromeric DNA of higher apes. We do, however, have a better understanding of the structure and role of the kinetochore. In all eukaryotes analyzed so far, the centromeres in a given genome separate in a sequential manner dependent upon the time of replication of pericentric and centromeric DNA. Some chromosomes, generally found in neoplastic cells, that carry more than one centromere show premature separation of the accessory centromeres. These centromeres and the associated pericentric regions replicate their DNA in an earlier part of the S phase than those that show kinetochore activity; both, however, carry DNA of the same composition. The active centromeres in these chromosomes show kinetochore protein binding as detected by antikinetochore antibody; the inactive centromeres are usually devoid of these proteins. The double minutes in neoplastic cells also lack kinetochore proteins, perhaps due to a lack of any centromere. Some dicentric and multicentric chromosomes in cancer cells and transformed cell lines do not display premature centromere separation. In these chromosomes, all centromeric sites show kinetochore proteins and all centromeric regions replicate their DNA simultaneously. These chromosomes also exhibited meiotic-like behavior of some centromeres and show postanaphase separation of some centromeres, resulting in bridges. These bridges, upon breakage and rejoining of sister chromatids, generate new multicentric chromosomes. The resulting chromosomes also exhibit formation of compound kinetochores. Some of these phenomena are novel descriptions of the centromere behavior in cancer cells. This review also discusses the role of aberrant centromere separation in human biology, providing correlates between errors of centromere separation and neoplasia.

Animals↗

Micronuclei, kinetochores and hypoploidy: tests with some agents.

Micronuclei were induced by treating mouse L-cells with diethylstilboestrol, colchicine and benomyl. These micronuclei were analysed for the presence of kinetochores by using antikinetochore antibody. The three chemicals induced micronuclei which differed with regards to their (i) relative distribution per cell, (ii) relative frequency for being kinetochore positive or being kinetochore negative and (iii) overall relationship between their induction and hypoploidy which in part may originate from laggards expressing as micronuclei. The data indicate that a study of micronuclei may help determine the differences between the actions of different chemicals on the genetic apparatus.

Alkylating Agents↗