Cell fusion studies and biochemical analysis of DNA synthesis in Werner and non-Werner cultured cells.
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Cell Fusion↗
Biomedical subjects
D Salk
Publications and source records attributed to D Salk.
Cytogenetic aspects of Werner syndrome.
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Chromosome Aberrations↗
Proteoglycans in the Werner syndrome and aging: a review and perspective.
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Aging↗
In vitro studies of Werner syndrome cells: aberrant growth and chromosome behavior.
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Cell Division↗
Induction of long-term immunity to paralytic poliomyelitis by use of non-infectious vaccine.
Naturally acquired immunity to paralytic poliomyelitis, which is of long duration, is associated with serum antibody, immunological memory, or both. Persistence of circulating antibody is reassuring but not essential for long-term protection. Immunological memory induced by killed poliovirus vaccine is similar to that induced by infection, which is not a prerequisite for the induction of long-term immunity. Animal studies of experimental vaccines indicate that some antigenic components of poliovirus induce immunological memory without producing detectable antibody; killed poliovirus vaccine has the same effect in man. Killed vaccine containing 40, 8, and 32 D-antigen units of poliovirus types 1, 2, and 3, respectively, protects all recipients in both one-dose and two-dose schedules. This means that either schedule can be chosen to fit in with local immunisation programmes and thus reduce costs and increase population coverage.
Antibodies, Viral↗
Flow cytometric analysis of small DNA content differences in heterogeneous cell populations: human amniotic fluid cells.
Accurate flow cytometric determination of nuclear DNA fluorescence has been attempted with human amniocentesis materials employing a detergent nuclear isolation protocol and 4,6-diamidino-2-phenylindole (DAPI) staining. Assay of native fluids yielded inadequate fluorescence histograms, whereas analysis of cultured amniocytes yielded unimodal histograms in 80 of 83 samples. Three fluids displayed unexpected bimodality, which may have resulted from the interaction between detergent and chromatin in rare cell types. Using a standardized protocol for preparation, staining, and assay, including brief periods of cocultivation of a human triploid reference standard, the percent experimental error in the flow cytometric DNA content estimate was as low as 0.2% in repeated assays of a single sample, and 0.4% in assays of a given genotype at different occasions. In two series of blindly assayed specimens, the percent standard error of the DNA content estimate ranged from 0.6 to 1.0%.
Amniotic Fluid↗
Clonal analysis of stable chromosome rearrangements in Bloom's syndrome fibroblasts.
In situ cytogenetic analysis was performed on colonies derived from single cells of cultured skin fibroblast-like strains from two patients with Bloom's syndrome (GM 1492 and GM 2520). Metaphases in all of the colonies displayed structural chromosome rearrangements. Among 212 metaphases from 24 colonies of GM 1492, only 16% were pseudodiploid, and there was a high incidence of de novo rearrangements within individual colonies. There were two "families" of 16 and five colonies, respectively, each containing identical or related aneusomies, and these could be arranged into pedigrees showing clonal evolution. The heterochromatic region of chromosome #1 and the telomeric regions of chromosome arms 2q, 3q, 4p, and 11q were most frequently involved in the rearrangements. In contrast, strain GM 2520 showed less intraclonal variation, was primarily pseudodiploid, and displayed only three clonal types, one of which had extensive subclonal variation (19 of 24 clones). A remarkable finding in GM 2520 was that, in some clones, extra copies of specific chromosome segments were present as translocations. These results caution against the use of strain GM 1492 as a prototype Bloom's syndrome strain for cell biological studies.
Bloom Syndrome↗
Vaccinology of poliomyelitis.
Vaccinology requires not only an understanding of the fundamental properties of the immune system and specific immunogens, but also the application of basic knowledge to develop effective immunization programmes for particular social, political, and environmental conditions. Two problems exist in the use of oral, live poliovirus vaccine: the occurrence of vaccine-associated paralytic poliomyelitis and the uncertain antibody response in tropical climates. These difficulties can be overcome by the use of killed poliovirus vaccine formulated to be effective with a single dose. Application of basic principles can bring poliomyelitis fully under control in all parts of the world.
Administration, Oral↗
Decreased oxygen supply enhances growth in culture of human mid-trimester amniotic fluid cells.
Human mid-trimester amniotic fluid cells were cultivated under conditions of decreased oxygen supply. Compared to control cultures the low-oxygen group showed improved growth which was quantitated by three independent assays (1) direct cell counts, (2) bromodeoxyuridine (BrdU)-Hoechst flow-cytometry, and (3) cloning efficiency. The growth promoting effects of lowered oxygen hold for all major morphologic categories of amniotic fluid cells.
Amniotic Fluid↗
Cultivated cells from mid-trimester amniotic fluids: IV. Cell type indentification via one and two-dimensional electrophoresis of clonal whole cell homogenates.
Clones of cultivated amniotic fluid cells that have distinct morphologic and growth characteristics (F, AF and E-type) were examined by one-dimensional SDS polyacrylamide gel electrophoresis (SDS-PAGE) and by two-dimensional electrophoresis employing isoelectric focusing and SDS-PAGE (IEF-PAGE). No qualitative differences in band pattern were observed in SDS-PAGE between the various amniotic fluid cell types, but consistent quantitative differences in the ratios of four bands of presumed filamentous proteins provided good distinction between amniotic fluid cells and postnatal skin fibroblast-like cells. By adding separation on the basis of electrical charge to that of molecular size (IEF-PAGE), we observed reproducible qualitative differences in the protein spot patterns between F and both AF and E-type amniotic fluid cells. At least eight discrete proteins appear not be synthesized by prenatal F-type cells in comparison with their isogenic AF and E counterparts under identical culture conditions. The two-dimensional electrophoretic patterns thus confirm that F and AF amniotic cells, in spite of their morphologic and growth kinetic similarities, are developmentally distinct cell types that retain their differentiated states in culture.
Amniotic Fluid↗
Werner's syndrome: a review of recent research with an analysis of connective tissue metabolism, growth control of cultured cells, and chromosomal aberrations.
Werner's syndrome is a rare, autosomal recessive condition with multiple progeroid features, but it is an imitation of aging rather than accelerated or premature senescence. Somatic chromosome aberrations occur in multiple tissues in vivo and in vitro, and there is an increased incidence of neoplasia. Thus. Werner's syndrome can be classified in the group of chromosome instability syndromes. Recent findings provide additional support for the concept that there is an aberration of connective tissue metabolism in Werner's syndrome, but it is unclear whether this is a primary or secondary manifestation of the underlying genetic defect. Abnormal growth characteristics are observed in cultured skin fibroblast-like cells and this provides another avenue for current research. Identification of the basic genetic defect in Werner's syndrome might clarify our understanding of the normal aging process in general, or might elucidate specific aspects such as the development of neoplasia, atherosclerosis, diabetes, or osteoporosis.
Aging↗
Morphological and biochemical heterogeneity of amniotic fluid cells in culture.
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Amniotic Fluid↗
Can we learn about aging from a study of Werner's syndrome?
Werner's syndrome, a rare genetic condition, has many features suggestive of rapid, premature aging. However, it differs from normal aging in several important respects. Cultured skin fibroblast-like cells have markedly reduced growth potential and a characteristic pattern of multiple, variable, stable structural chromosomal rearrangements that are non-constitutional. Since Werner's syndrome is inherited in an autosomal recessive fashion, there is presumably a single gene defect or a multi-gene deletion that accounts for its multiple progeroid features. The identification of this defective protein (or proteins) would be of great interest to both basic gerontologic and clinical geriatric research, since it would shed light on the mechanisms underlying the normal aging process and the development of geriatric diseases. It is evident that such a protein(s) is not the sole determinant of youth or longevity, but the many progeroid manifestations of Werner's syndrome indicate that the basic molecular defect resides in or affects some central pathway of cellular control or homeostatic interaction.
Adolescent↗
Effects of radical-scavenging enzymes and reduced oxygen exposure on growth and chromosome abnormalities of Werner syndrome cultured skin fibroblasts.
Two sister strains of skin fibroblast-like (FL) cells from a patient with Werner's syndrome (adult progeria) were grown in regular tissue culture medium or medium supplemented with the radical-scavenging enzymes superoxide dismutase and catalase. Differences in proliferative potential were shown to be due to strain variation rather than to treatment with the enzymes. Two independent strains were also grown in ambient (18%) or reduced (1%) concentrations of oxygen. All cultures (untreated, treated with enzymes, treated with reduced oxygen exposure) displayed the limited in vitro growth potential and cytogenetic abnormality characteristic of Werner's syndrome FL cells; thus the underlying defect in Werner's syndrome does not appear to be related to defective protection against or repair of damage by free radicals.
Catalase↗
Systematic growth studies, cocultivation, and cell hybridization studies of Werner syndrome cultured skin fibroblasts.
The growth of 20 independently derived skin fibroblastlike (FL) cell strains from three individuals with Werner syndrome (adult progeria) was compared with the growth of ten FL cell strains from normal individuals. Population growth rates and total replicative life spans of Werner syndrome strains averaged 53% and 27%, respectively, of the growth rates and life spans of non-Werner strains. In the first few passages, four Werner syndrome strains demonstrated population growth rates in the low normal range, but the longest-lived Werner syndrome strain had only 75% of the total replicative potential of the shortest-lived normal strain. Exponential growth rates, cloning efficiencies, and saturation densities of Werner strains were also reduced, whereas cell attachment was normal. Viable cells (identified by dye exclusion) were maintained in post-replicative Werner syndrome and control cultures for periods of at least 10 months; there was no evidence of accelerated post-replicative senescence of cell death of Werner syndrome FL cells. Cocultivation of Werner syndrome and normal strains did not influence population growth rates of either strain. Two proliferating hybrid clones were obtained from fusions of normal and Werner syndrome FL cell strains and these hybrids displayed the reduced growth potential typical of Werner syndrome FL cells. These studies confirm that low growth rate and sharply reduced replicative life span are characteristic of cultured skin FL cells from patients with Werner syndrome, and they suggest that these characteristics are not affected by complementation with non-Werner FL cells.
Cell Division↗
Antenatal diagnosis of mosaic trisomy 8 confirmed in fetal tissues.
An aneuploid karyotype with an extra submetacentric C-group chromosome was observed in all metaphase cells in 5 of 24 primary amniotic fluid cell clones and in admixture with normal cells in two additional clones. Trisomy 8 was demonstrated by R-banding. The parents elected to terminate the pregnancy. Successful cultures were initiated from nine fetal tissues and aneuploid cells were observed in cultures derived from two separate sites in the skin.
Adult↗
Cytogenetics of Werner's syndrome cultured skin fibroblasts: variegated translocation mosaicism.
Skin fibroblast-like (FL) cells from patients with Werner's syndrome (adult progeria) regularly demonstrate frequent pseudodiploidy involving variable structural rearrangements that are clonal: variegated translocation mosaicism (VTM). Ninety-two percent of 1,538 metaphases from 29 independent strains derived from five patients with Werner's syndrome demonstrated this cytogenetic abnormality. In contrast, only eight (8.4%) of 95 non-Werner's syndrome FL cell cultures demonstrated VTM: seven with low-grade VTM (approximately 5% of 300 metaphases), and one with VTM affecting 90-100% of metaphases. Unlike the cytogenetic abnormalities observed in the terminal stages of normal FL cell cultures, VTM occurs throughout the entire lifespan of Werner's syndrome cultures. Ten of the identifiable break points in 1,005 banded metaphases accounted for 27% of all definable rearrangements. Baseline sister chromatid exchanges were not increased. Cocultivation of Werner's syndrome and normal strains did not induce VTM in the normal strain. The relationship between VTM and the reduced growth potential of Werner's syndrome FL cells is not yet understood, nor is the relationship between these in vitro abnormalities and the presumptive single gene defect that causes the progeroid clinical manifestations of Werner's syndrome.
Adolescent↗