Cloning of embryonal stem cell-specific genes: characterization of the transcriptionally controlled gene esg-1.

We have isolated, by differential library screening, eight cDNAs representing genes that are specifically expressed in the embryonal stem cell line IMT-11, when compared to the parietal endoderm-like cell line PYS-2 or to NIH3T3 fibroblasts. One of these genes, embryonal stem cell gene 1 (esg-1), was analyzed in detail. esg-1 mRNA is found at high levels in both IMT-11 and F9 embryonal carcinoma cells and disappears during the differentiation of the stem cells. Furthermore, expression of the gene was found to be extremely low in, or absent from, oocytes and fertilized eggs, but it is strongly induced at the 2-cell stage, reaching maximum levels at the 4-cell stage. In contrast, esg-1 expression is detectable neither in midgestation embryos nor in neonatal tissues. These results strongly suggest that esg-1 is expressed specifically or at least predominantly in embryonal stem cells. Antibodies directed against a glutathione S-transferase-esg-1 fusion product detect a protein of M(r) approximately 14,000 in F9 embryonal carcinoma cells, but not in differentiated cells. Apart from the esg-1 gene, which contains two introns, there are at least seven esg-1-related pseudogenes in the mouse genome that differ from the esg-1 gene by the presence of multiple point mutations, by the lack of intervening sequences, and/or by the presence of a polyadenylated stretch at the 3' end. The esg-1 gene is under stringent transcriptional control in differentiating and differentiated cells, as shown by both nuclear run-on assays and the transient F9 stem cell-specific expression of constructs consisting of esg-1 upstream sequences fused to a luciferase reporter gene.

Macronuclear and micronuclear configurations of a gene encoding the protein synthesis elongation factor EF 1 alpha in Stylonychia lemnae.

The micronuclear and macronuclear configurations of a gene encoding the protein synthesis elongation factor EF 1 alpha in the hypotrich ciliate Stylonychia lemnae were compared. The two sequences are generally colinear. The coding sequence of the micronuclear gene is, however, interrupted by a 64 bp insert flanked by a 2 bp direct repeat in a gene region which is moderately conserved among EF 1 alpha genes of different organisms. The insertion site is distinct from known intron positions in eukaryotic EF 1 alpha genes. The insert sequence shows inverted repeats at its ends and thus exhibits typical features of an internal eliminated sequence (IES). Comparison with other such sequences in the related organism Oyxtricha nova shows that the IES falls into a new group of such elements. The macronuclear gene exhibits a strikingly limited codon usage, which cannot be simply explained by the overall base composition of the DNA but probably also relates to the very high copy number of the macronuclear gene and the putative high amount of the gene product.

Case-control study of lung cancer in civilian employees at the Portsmouth Naval Shipyard, Kittery, Maine.

Case-control analysis of deaths due to lung cancer (International Classification of Diseases, Eighth Revision, code 162) among persons who worked at the Portsmouth Naval Shipyard, Kittery, Maine, between 1952 and 1977 found elevated odds ratios for exposures to ionizing radiation, asbestos, and welding byproducts. The radiation-related excess was statistically significant in persons with cumulative lifetime exposures of 1.0-4.999 rem. When asbestos and welding histories were combined into a single risk factor, odds ratios for the combined exposure were significantly elevated for two of three duration-of-exposure categories examined. Further analysis of data on radiation exposure, controlling for exposures to asbestos and welding, found reductions in initial estimates of radiation risk at all levels of radiation exposure. This reduction suggests that radiation workers were more heavily exposed to asbestos and/or welding fumes than were other workers and that those exposures confounded the observed association between radiation and lung cancer. Analysis of mortality by time since first exposure to radiation revealed no pattern of progressive increase as latency increased. By contrast, odds ratios for asbestos/welding increased with latency. Data on cigarette smoking and socioeconomic status were not available. The results of this study do not preclude a possible association between radiation exposure at the Portsmouth Naval Shipyard and excess mortality from lung cancer. However, they provide no evidence in support of such a relation.

A case-control study of leukemia at a naval nuclear shipyard.

A matched case-control study was conducted of 53 leukemia deaths and of 212 controls within a previously studied cohort of 24,545 on-shore workers employed between January 1, 1952 and August 15, 1977 at the Portsmouth (New Hampshire) Naval Shipyard. The study sought to ascertain a priori whether there was an association between leukemia deaths and occupational exposure to either ionizing radiation or organic solvents. To obtain information on individual exposures, radiation dose histories and detailed work histories by job and shop were evaluated for each subject. No statistically significant associations were found either between ionizing radiation or presumed solvent exposure and myelogenous or lymphatic leukemia. However, when specific job categories and shops were examined without benefit of a priori hypotheses, two occupations, electrician and welder, were found to be associated with leukemia. For electricians, the Mantel-Haenszel odds ratio (ORMH) was significantly elevated for all leukemias (ORMH = 3.00, 95% confidence interval (CI) = 1.29-6.98), particularly for lymphatic leukemia (ORMH = 6.00, 95% CI = 1.47-24.45). For welders, the odds ratio was not significantly elevated for all leukemias (ORMH = 2.25, 95% CI = 0.92-5.53), but was significantly elevated for myeloid leukemia (ORMH = 3.83, 95% CI = 1.28-11.46). These findings persisted when potential confounders were adjusted by means of a conditional logistic regression model.

Cancer mortality at a Naval Nuclear Shipyard.

To evaluate a reported five-fold increase in leukaemia mortality among workers exposed to ionising radiation at Portsmouth (New Hampshire) Naval Shipyard (PNS), a retrospective cohort mortality study of all PNS civilian workers employed from 1952 to 1977 was done. Three subcohorts were identified: 7615 workers with radiation exposure of 0.001 to 91.414 rem (mean 2.779 rem, median 0.545 rem), 15 585 non-radiation workers, and 1345 workers selected for radiation work who received no measurable exposures. Vital status on 96% of the workers was ascertained and observed mortality due to all causes, all malignant neoplasms, and malignant neoplasms of the lymphatic and haematopoietic tissues, including leukaemia, was compared with that expected from mortality-rates for United States White males. Leukaemia mortality in radiation and non-radiation workers at PNS was also compared. Although the study had a power of greater than 99% to detect statistically a five-fold increase in leukaemia mortality among the radiation workers, and a power of 67% to detect a two-fold increase, there was no excess due to leukaemia or any other cause. The standardised mortality ratio for leukaemia among radiation workers was 84 (95% confidence interval, 34--174). There was no dose-response relation with radiation or any increased mortality in radiation over non-radiation workers. The study was, however, limited by short latency (time since first radiation); only 53% of the workers had less than 15 years' latency.

Technological feasibility of the 2 fibers/cc asbestos standard in asbestos textile facilities.

The technical feasibility of developing dust control practice which would bring the asbestos industry into compliance with the OSHA I fibers/cc standard, scheduled to become effective July 1, 1976, has been questioned since its promulgation. This paper presents the results of industrial hygiene surveys, including the results of 243 asbestos air samples, conducted at two asbestos textile production facilities which have achieved the 2 fibers/cc standard. Both facilities use nonconventional production techniques. Conclusions drawn from the investigations express the need for: (1) a study of new products; (2) a tool to motivate consumers to switch to the new products when appropriate; and (3) legislation which will protect all domestic producers of asbestos textiles from certain producers who do not have to absorb the added expense of meeting the OSHA standard.