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The effect of radiation-induced mutations on the fitness of Drosophila populations.

The change in frequencies of D. melanogaster and D. simulans in competition experiments was used to measure the effect of radiation on the fitness of a population. A dose of 250 or 500 rads given to the males of highly inbred lines of D. simulans at the beginning of competition and every three weeks thereafter increased the relative frequency of the irradiated population. If the dose was increased to 1000 rads, the deleterious effects of radiation became too great a burden on the population, and the frequency of the irradiated population decreased. From these results it was concluded that below certain doses the introduction of radiation-induced mutations into a highly homozygous population would increase the fitness of the population.

Animals↗

A simple murine model for immobilization osteopenia.

Reduction of loading force to bone induces osteopenia. Although the tail suspension model is the frequently used osteopenia model, this model burdens the animals with nonphysiologic blood distribution and systemic stress. We developed a new simple animal model for osteopenia under reduced loading. Both hind legs of male ICR mice (8 weeks old) in the experimental group were inserted into plastic tubes, which then were connected with wires of three sizes. This apparatus completely immobilized both femurs while allowing the tibias to move. Animals were pair-fed and sacrificed on Days 3, 7, 10, and 14 after immobilization. Bone mineral density measurement with dual energy xray absorptiometry revealed bone loss in the immobilized femurs. Histomorphometric analysis showed increased bone resorption and decreased bone formation, which started from Day 7 and continued until Day 14, resulting in structural disorders in the cancellous bone. Osteoclast population increase before osteoblast population decrease revealed that osteoclasts initially affect the process of this type of osteopenia. Our immobilization model is simple, easy to use, well-tolerated by the animal, and has potential for evaluating the therapeutic effects of drugs to treat osteopenia caused by reduced loading.

Animals↗

Variation in heterotrophic and autotrophic nitrifier populations in relation to nitrification in organic soils.

The occurrence of heterotrophic and autotrophic nitrifiers in Pahokee muck and the role of these organisms in the ecosystem were assessed by surveying their population densities under different field conditions and by observing the relationship of these populations with aerobic bacteria and soil moisture. Heterotrophic nitrifier populations varied from 2.0 x 10 to 3.8 x 10 bacteria per cm of muck in surface fallow (bare) Pahokee muck during the annual cycle. This population decreased 40-fold between the surface and the 60- to 70-cm depths of soil. Similar variations were noted with autotrophic nitrifier populations. Significant correlations were found between heterotrophic nitrifiers and both soil moisture and aerobic bacteria. These relationships did not exist for the autotrophic nitrifiers. In soil that had been heated to kill the autotrophic nitrifiers, while preserving a population of the heterotrophs, and then amended with sodium acetate or ammonium sulfate or both, no nitrate or nitrite accumulated, although significant increases in heterotrophic nitrifiers were detected. In unheated control soil, nitrate plus nitrite-N increased from 14.3 to 181 mug/g of wet soil, and 48 mug of nitrite-N per g was produced. These data suggest that the autotrophic nitrifiers were the sole population responsible for nitrification in Pahokee muck.

Journal Article↗

Apolipoprotein E phenotypes in Finnish youths: a cross-sectional and 6-year follow-up study.

Apolipoprotein E (apoE) polymorphism is a genetic determinant of plasma lipid levels and of coronary heart disease (CHD) risk. We determined the apoE phenotypes and plasma lipid levels in 1577 youths aged 3 to 18 years in 1980. The subjects were randomly selected from five areas of Finland. ApoE phenotyping was performed directly from plasma by isoelectric focusing and immunoblotting. The apoE allele frequencies in the population sample were epsilon 2 = 0.039, epsilon 3 = 0.767, and epsilon 4 = 0.194. There were no differences in the apoE phenotype distribution between East and West Finland or between sexes. The concentrations of serum total cholesterol, low density lipoprotein cholesterol, and apolipoprotein B increased with apoE phenotype in the order of E2/2, E3/2, E4/2, E3/3, E4/3, and E4/4. This increase was already seen in 3-year-old children; it was observed in both sexes, but was clearer in males than in females. The mean levels of high density lipoprotein (HDL) cholesterol, apolipoprotein A-I, triglyceride, Lp[a] lipoprotein, and the activity of lecithin:cholesterol acyltransferase did not differ between the apoE phenotypes. The observed differences in serum cholesterol remained fairly stable during the 6-year follow-up from 1980 to 1986, while the mean serum cholesterol concentration in the whole study population decreased by 6.3%. This study confirms the reported higher frequency of the epsilon 4 allele in Finns as compared to most other populations; this may contribute to the high rates of CHD in Finland as compared to most other populations. The results do not, however, explain the higher rate of CHD in East Finland in comparison to the western part of the country.

Adolescent↗

Population size and the nature of genetic load in Gentianella germanica.

Theory predicts a significant relationship between the size of a population and the magnitude and composition of its genetic load, but few natural populations have been investigated. We examined the magnitude of genetic load due to recessive deleterious alleles (GL) both segregating and fixed within Gentianella germanica populations of varying size by selfing and reciprocally crossing plants within and between natural populations according to a partial diallel design and by comparing the performance of the experimental progeny in a common-garden experiment. The results show that GL for total fitness in small populations (fewer than 200 plants) was mainly due to fixed recessive deleterious alleles, whereas GL for total fitness in larger populations (more than 200 plants) appeared to be mainly due to segregating deleterious recessive alleles. The total fitness of selfed plants increased with decreasing population size, indicating some purging of deleterious alleles associated with declining population sizes. The magnitudes of GL due to fixed deleterious alleles in small populations and segregating deleterious alleles in large populations, however, were overall similar, suggesting that purging selection was an insignificant force when compared to genetic drift in determining the magnitude of GL in small natural populations in this species. The results of this study highlight the importance of population size in determining the dynamics of genetic loads of natural populations and are overall in line with a large body of theoretical work indicating that small populations may face higher extinction risks due to the fixation and accumulation of deleterious alleles of small effect.

Alleles↗

Isolation and functional characterization of human intestinal mucosal lymphoid cells.

Viable suspensions of human colonic mucosal lymphoid cells have been prepared by sequential treatment of tissue with dithiothreitol, EDTA in calcium- and magnesium-free salt solutions, and purified collagenase. The intestinal lymphocyte population, in comparison with that of peripheral blood, had greater numbers of bone marrow-derived cells, particularly cells bearing membrane IgA; showed spontaneous association with macrophages; underwent rapid rosette formation with sheep erythrocytes; and demonstrated increased in vitro synthesis of immunoglobulin. Total thymus-derived cells were equal in the two populations. Decreases were found in "null" cell numbers, in cells bearing membrane IgD and IgM, and in responsiveness to phytohemagglutinin. Macrophage/monocytes in the intestinal population were increased in size, granularity, motility, sustained glass adherence, and phagocytic activity. Human intestinal lymphoid cells appear to constitute a cell population that is more "mature" and/or "activated", in comparison with the lymphoid cells of peripheral blood. The method of preparation should lend itself to the study of inflammatory bowel disease, gastrointestinal cancer, and the intestinal secretory immune system.

Antibody Formation↗

Relaxin production and release after hysterectomy in the pig.

Relaxin secretion by luteal tissue and into peripheral blood was examined in relation to changes in the fine structure of aging porcine corpora lutea. Sequential bleedings every sixth day reveal a similar increase in serum relaxin concentrations from days 6-114 in hysterectomized and pregnant gifts. The span of the estrous cycle is about 21 days, and the length of pregnancy is about 115 days in pigs. Estrone (E1) and 17 beta-estradiol (17 beta-E2) serum levels increase with placental development; after hysterectomy, they are low from days 12-168. In luteal tissue, peak quantities of relaxin and electron-dense granules occur on day 100 in hysterectomized and pregnant gilts. By day 112, luteal tissue relaxin levels and granule populations decrease by about half, while relaxin secretion into peripheral blood approaches peak concentrations about day 114 in hysterectomized as well as pregnant gilts. Between days 114 and 120, serum relaxin decreases by half even though the corpora lutea persist to day 150 in hysterectomized animals. Thereafter, cytoplasmic granules and blood and luteal tissue levels of relaxin decrease gradually to day 150 in hysterectomized gilts; in contrast, they disappear abruptly after parturition. In hysterectomized gilts, relaxin release coincides with diminished populations of granules in corpora lutea, and the initiation of these events seems to be precisely timed. E1 and 17 beta-E2 secretion is unrelated to relaxin secretion in pregnant and hysterectomized gilts. High peripheral blood levels of E1 and 17 beta-E2, primarily of placental origin, or even the presence of the uterus are not required in the release of relaxin at day 114. By using steroidogenic and protein synthetic capabilities of luteal cells as criteria essential to the evolution of reproduction in this species, the results presented here suggest that 114 days rather than 21 days characterize the reproductive cycle, even in the absence of the uterus. These results indicate that the production and, particularly, the release of relaxin into peripheral blood on about day 114 in hysterectomized gilts as well as pregnant animals may be genetically controlled.

Animals↗

Effects of in-situ ozonation on indigenous microorganisms in diesel contaminated soil: survival and regrowth.

Soil column experiments were conducted to investigate the effects of chemical oxidation on the survival of indigenous microbes (i.e., heterotrophic microbes, phenanthrene-degrading microbes, and alkane-degrading microbes) for field soil contaminated with diesel fuel. Rapid decreases of total petroleum hydrocarbons (TPH) and aromatics of diesel fuel were observed within the first 60 min of ozone injection; after 60 min, TPH and aromatics decreased asymptotically with ozonation time. The three types of indigenous microbes treated were very sensitive to ozone in the soil column experiment, hence the microbial population decreased exponentially with ozonation time. The numbers of heterotrophic, alkane-degrading, and phenanthrene-degrading bacteria were reduced from 10(8) to 10(4), 10(7) to 10(3), and 10(6) CFU g soil(-1) to below detection limit after 900 min of ozonation, respectively. Except for the soil sample ozonated for 900 min, incubation of ozone-treated soil samples that were not limited by oxygen diffusion showed further removal of TPH. The soil samples that were ozonated for 180 min exhibited the lowest concentration of TPH and the highest regrowth rate of the heterotrophic and alkane-degrading populations after the 9 weeks of incubation.

Bacteria↗

The interaction among evolutionary forces in the pathogenic fungus Mycosphaerella graminicola.

The population genetic dynamic of a species is driven by interactions among mutation, migration, drift, mating system, and selection, but it is rare to have sufficient empirical data to estimate values for all of these forces and to allow comparison of the relative magnitudes of these evolutionary forces. We combined data from a mark-release-recapture experiment, extensive population surveys, and computer simulations to evaluate interactions among these evolutionary forces in the pathogenic fungus Mycosphaerella graminicola. The results from these studies showed that, on average, the immigration rate was 0.027, the fraction of outcrossing individuals was 0.035, and the selection coefficient associated with immigrants was 0.106 each generation. We also estimated that effective population sizes for this fungus were larger than 24,000 and the mutation rate for the RFLP markers used in surveys and field experiments was approximately 4 x 10(-5). Computer simulations based on these estimates indicate that, on average, the global population of M. graminicola has reached equilibrium. Population genetic parameters including number of alleles, gene diversity, and population subdivision estimated from the computer simulations were surprisingly close to empirical estimates. Simulations also revealed that random drift is the major evolutionary force decreasing genetic variation in this fungus, followed by natural selection. The major force adding to genetic variation was mutation, followed by gene flow and sexual recombination. Gene flow played the leading role in decreasing population subdivision while natural selection was the major factor increasing population subdivision.

Ascomycota↗

Long-term services in Mexico: homes for the aged.

Accurate statistics are not available about the number of people in institutions for the aged in Mexico, but the majority are assumed to live with their family or on their own and to be integrated with the community. However, the increase of Mexico's aging population, from its present more than 4 million to 12 million people over 60 years of age in the year 2025, together with the decreased population growth rate (3.5% in the early seventies; approximately 2.5% currently, and 1% in the year 2000) will considerably increase the demand for health care and services for this sector of the population. Nursing Homes should be not only shelters, but should help to raise or maintain the quality for life of elderly people in all areas: physical, psychological, and social. In the near future, it will be necessary not only to build more homes for the aged, but to develop and implement other forms of care that will allow the elderly to remain integrated with the community. Through home care and substitute homes, families that take care of their elderly relatives can be helped; and direct assistance can be give to old people who wish to be independent.

Aged↗

An evaluation of the decreasing incidence of positive surgical margins in a large retropubic prostatectomy series.

PURPOSE: Positive surgical margins adversely affect biochemical recurrence-free survival after radical retropubic prostatectomy (RRP) for prostate cancer. We retrospectively reviewed a large series of men who underwent RRP at a single academic university urology program to define the change in the incidence of organ confined (OC) disease and positive surgical margin (SM+) during the last 2 decades. MATERIALS AND METHODS: Between 1982 and 2001, 9,035 men underwent RRP for clinically localized prostate cancer (T1 to T3a) at a single institution. We compared the incidences of OC disease and SM+ in this population. RESULTS: An increasing proportion of men presented with OC disease over time. The incidence of SM+ in the overall RRP population decreased dramatically over time. However, in men with nonorgan confined disease (pT3), the proportion with SM+ was stable and consistently elevated (22.7% to 27.8%), after the initial decrease from the early 1980s (53%). CONCLUSIONS: Widespread early detection programs for prostate cancer resulted in a downward stage migration in men presenting with clinically localized prostate cancer at our institution during the last 2 decades. The decrease in the percentage of men with SM+ was due to the increasing number of men with organ confined disease. These results imply that the decrease in surgical margin rates in the overall RRP population is most likely due to stage migration and improved patient selection, rather than major improvements in surgical technique. The stable percentage of SM+ among men with pT3 disease dictates the need for continued evaluation of surgical technique and the need for effective adjuvant therapy.

Adenocarcinoma↗

[Obesity and diabetes-morbidity--a prospective 5-year study].

In a prospective 5-year-study the development of adiposity was tested in a population group consisting of about 30,000 persons. It was shown that during the time of investigation the population decreased in its total number, the consumption of easily digestible carbohydrates and meat, however, increased. As it was established in 1968, the proportion of the adipose persons (more than 20% of the Broca-weight) and the percental average deviation from the Broca-weight (calculated in decennium classes) was more than in the comparable population groups in the GDR. Contrary to expectation the examination further showed that on an average men exhibited a trend to the increase in weight, but on the average women decreased in weight. In all age groups (with the exception of the 60- to 69-year-old persons) the proportion of obese persons was significantly lower in 1973 than in 1968. Unchangedly adipose are the diabetics who also have a by far higher average weight than the healthy population and who up to now do not reveal an inclination to the decrease in weight. With 2.9% the average frequency of diabetes is also adequately high.

Adult↗

Recovery of HeLa cell population growth after treatment with 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB).

Dose-response curves for the inhibition of heterogeneous nuclear RNA (hnRNA) synthesis in HeLa cells by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB; 5-100 microM; 30 min) are biphasic and indicate the existence of two subpopulations of hnRNA molecules, one highly sensitive and the other completely resistant, as previously reported for molecules greater than 1,000 nucleotides long (Tamm et al., 1976; Sehgal et al., 1976a). In the short-term experiments, the drug-sensitive synthesis of hnRNA was inhibited 50% at a DRB concentration of approximately 7 microM, and 70% at 20 microM, whereas drug-resistant synthesis, which comprises approximately 20% of total, continued at DRB concentrations as high as 100 microM. After 24 hr of DRB treatment in medium containing 5% fetal calf serum, the increase in cell number in the exponentially growing population was inhibited by only 42% at 20 microM DRB, and the formation of colonies of greater than or equal to ten cells was not decreased. DRB at 40 microM concentration decreased population growth by 76% and colony formation by 63%. Treatment with 60 microM DRB was sufficient to prevent a net increase in cell number and to reduce colony formation by 78%. After termination of treatment, the time required for the surviving population to begin rapid proliferation was directly related to the concentration of DRB used to treat cells and to the duration of treatment. After 24-hr treatment with 40 microM DRB, cultures recovered within 1 day, whereas after 60 microM DRB, 3-4 days were required. After 40-hr treatment with 60 microM DRB, 5-6 days were required for recovery, and after 80 microM DRB, 9-11 days. During the "dormant" period the cell number ranged from 15 to 60% of the initial number and was fairly stable for given conditions. After the "dormant" period, recovery was rapid. The population growth rate in cultures undergoing treatment with DRB is directly related to serum concentration; however, the recovery rate during the post-treatment period is unaffected by serum concentration.

Blood↗

Microbial characterization during composting of municipal solid waste.

This study investigates the prevailing physico-chemical conditions and microbial community; mesophilic bacteria, yeasts and filamentous fungi, bacterial spores, Salmonella and Shigella as well as faecal indicator bacteria: total coliforms, faecal coliforms and faecal Streptococci, present in a compost of municipal solid waste. Investigations were conducted in a semi-industrial pilot plant using a moderate aeration during the composting process. Our results showed that: (i) auto-sterilization induced by relatively high temperatures (60-55 degrees C) caused a significant change in bacterial communities. For instance, Escherichia coli and faecal Streptococci populations decreased, respectively, from 2 x 10(7) to 3.1 x 10(3) and 10(7) to 1.5 x 10(3) cells/g waste dry weight (WDW); yeasts and filamentous fungi decreased from 4.5 x 10(6) to 2.6 x 10(3) cells/g WDW and mesophilic bacteria were reduced from 5.8 x 10(9) to 1.8 x 10(7) bacteria/g WDW. On the other hand, the number of bacterial spores increased at the beginning of the composting process, but after the third week their number decreased notably; (ii) Salmonella disappeared completely from compost by the 25th day as soon as the temperature reached 60 degrees C; and (iii) the bacterial population increased gradually during the cooling phase. While Staphylococci seemed to be the dominant bacteria during the mesophilic phase and at the beginning of the thermophilic phase, bacilli predominated during the remainder of the composting cycle. The appearance of gram-negative rods (opportunistic pathogens) during the cooling phase may represent a serious risk for the sanitary quality of the finished product intended for agronomic reuse. Compost sonication for about 3 min induced the inactivation of delicate bacteria, in particular gram-negatives. By contrast, gram-positive bacteria, especially micrococcus, spores of bacilli, and fungal propagules survived, and reached high concentrations in the compost.

Biodegradation, Environmental↗

Evolution of reduced pre-adult viability and larval growth rate in laboratory populations of Drosophila melanogaster selected for shorter development time.

Four large (n > 1000) populations of Drosophila melanogaster, derived from control populations maintained on a 3 week discrete generation cycle, were subjected to selection for fast development and early reproduction. Egg to eclosion survivorship and development time and dry weight at eclosion were monitored every 10 generations. Over 70 generations of selection, development time in the selected populations decreased by approximately 36 h relative to controls, a 20% decline. The difference in male and female development time was also reduced in the selected populations. Flies from the selected populations were increasingly lighter at eclosion than controls, with the reduction in dry weight at eclosion over 70 generations of selection being approximately 45% in males and 39% in females. Larval growth rate (dry weight at eclosion/development time) was also reduced in the selected lines over 70 generations, relative to controls, by approximately 32% in males and 24% in females. However, part of this relative reduction was due to an increase in growth rate of the controls populations, presumably an expression of adaptation to conditions in our laboratory. After 50 generations of selection had elapsed, a considerable and increasing pre-adult viability cost to faster development became apparent, with viability in the selected populations being about 22% less than that of controls at generation 70 of selection.

Adaptation, Physiological↗

Population differentiation in an annual legume: genetic architecture.

The presence or absence of epistasis, or gene interaction, is explicitly assumed in many evolutionary models. Although many empirical studies have documented a role of epistasis in population divergence under laboratory conditions, there have been very few attempts at quantifying epistasis in the native environment where natural selection is expected to act. In addition, we have little understanding of the frequency with which epistasis contributes to the evolution of natural populations. In this study we used a quantitative genetic design to quantify the contribution of epistasis to population divergence for fitness components of a native annual legume, Chamaecrista fasciculata. The design incorporated the contrast of performance of F2 and F3 segregating progeny of 18 interpopulation crosses with the F1 and their parents. Crosses were conducted between populations from 100 m to 2000 km apart. All generations were grown for two seasons in the natural environment of one of the parents. The F1 often outperformed the parents. This F1 heterosis reveals population structure and suggests that drift is a major contributor to population differentiation. The F2 generation demonstrated that combining genes from different populations can sometimes have unexpected positive effects. However, the F3 performance indicated that combining genes from different populations decreased vigor beyond that due to the expected loss of heterozygosity. Combined with previous data, our results suggest that both selection and drift contribute to population differentiation that is based on epistatic genetic divergence. Because only the F3 consistently expressed hybrid breakdown, we conclude that the epistasis documented in our study reflects interactions among linked loci.

Biological Evolution↗

Impact of dried skim milk in production diets on Lactobacillus and pathogenic bacterial shedding in growing-finishing swine.

AIMS: To determine the possible effects of inclusion of dried skim milk (DSM) in swine diets on indigenous Lactobacillus spp. and Escherichia coli, and its potential for controlling pathogen shedding and affect animal growth in growing-finishing swine. METHODS AND RESULTS: Animals were fed over three dietary phases to match production needs from age 10-14 weeks, 14-18 weeks and 18-22 weeks. For each feeding phase, diets were formulated to contain 0 or 10% DSM (balanced for metabolizable energy and true ileal digestible amino acids). Animals were weighed every 2 weeks and faecal samples were collected from 40 animals (20 with DSM and 20 without DSM) at week 10 (d 0 on diets), 14, 18 and 22 of age, and were analysed for Lactobacillus spp., Enterobacteriaceae, coliforms, E. coli, Salmonella, Campylobacter and E. coli O157:H7. At the start of the study (week 10), faecal bacterial counts (log10 CFU g(-1) faeces) were 9.55, 7.26, 7.01 and 6.93 for Lactobacillus, Enterobacteriaceae, coliforms and E. coli populations respectively. The Enterobacteriaceae, coliform and E. coli populations decreased through week 14 and 18, but were higher in animals fed with the DSM diet compared with the basal diet without DSM. The Lactobacillus populations at weeks 14 and 18 were lower in the animals fed the diet without DSM, whereas feeding DSM maintained the Lactobacillus counts from week 10. At week 22, populations of Enterobacteriaceae, coliforms and E. coli were >week 18 for the animals fed the diet without DSM, less change was observed with the feeding of DSM, and no differences between the diets were observed at week 22. However, in week 22 the animal gain was positively correlated with Lactobacillus numbers and negatively correlated with E. coli numbers. Subtraction of the E. coli population (log10) from the Lactobacillus population (log10) yielded a positive value termed 'effective'Lactobacillus that correlated well with animal gain and may better define a beneficial function in the intestine. Salmonella were detected in over 60% of the animals at week 10 and 14, and <20% at week 18 and 22. Campylobacter were detected rarely at weeks 10, 14 and 18, but were found in 25% of the animals at week 22. The DSM did not affect Salmonella or Campylobacter shedding, but examination of individual animals over the entire experiment indicated that fewer recurring incidences of Salmonella shedding occurred in animals that maintained higher Lactobacillus. In addition, at week 22, Salmonella and Campylobacter shedding was associated with lower levels of effective Lactobacillus and lower animal weight gains. CONCLUSIONS: The DSM did not directly affect the animal performance or pathogen shedding via the Lactobacillus spp. population at any phase of production. However, analysis of data from all animals revealed that faecal Lactobacillus affected Salmonella shedding and in the finishing phase, animal growth and pathogen shedding also were affected, as reflected by the 'effective'Lactobacillus-associated observations. SIGNIFICANCE AND IMPACT OF THE STUDY: In the swine intestine, any benefits from gastrointestinal Lactobacillus may be compromized by the E. coli population, and this antagonism may explain responses observed with prebiotics or probiotics in some swine.

Animal Feed↗

Inhibition of DNA synthesis by a small-cell lung carcinoma-derived protein.

A tumor-derived factor that inhibits cellular DNA synthesis was identified. The factor was extractable from a small-cell lung carcinoma cell line grown in either chemically defined medium or nu/nu mice and inhibited tritiated thymidine ([3H]dThd) incorporation by tumor cell lines of autologous, allogeneic, and xenogeneic origins. The viability of nonproliferating cells from normal tissue was not affected. Tumor extract inhibitory activity was trypsin labile but was resistant to other proteases, neuraminidase, lipase, DNase, RNase, glucosidase, extremes of pH-temperature, and reducing conditions. Inhibitory activity was reversibly bound to helix pomatia lectin but not to lentil, wheat germ, or concanavalin A lectins. Purification by size-exclusion high-performance liquid chromatography yielded a bioactive unimodal 12-kilodalton (kd) peak. The bioactive 12-kd moiety could be eluted from sodium dodecyl sulfate-polyacrylamide gels. Redosing of populations of the T-lymphoblastoid cell line CEM achieved an early (24 hr) sustained depression of pulse [3H]dThd incorporation and ultimately led to decreased population density of factor-treated populations. DNA histogram analysis demonstrated no change in cell cycle phase distribution after factor treatment. 5-Bromo-2'-deoxyuridine (BrdUrd) vs. propidium iodide with the two-parameter Fluorescence-Activated Cell Sorter analysis showed relative inhibition of non-S-phase BrdUrd uptake at 24 hours. A cell-free DNA polymerase assay demonstrated significant inhibition of non-alpha-polymerase-associated DNA synthesis in factor-treated cells. These studies suggest that this tumor-derived inhibitor of DNA synthesis represents a class of cellular products involved in the autoregulation of growth by regulation of DNA synthetic activity.

Bromodeoxyuridine↗