Bcl-2 inhibits glucocorticoid-induced apoptosis but only partially blocks calcium ionophore or cycloheximide-regulated apoptosis in S49 cells.

Many non-Hodgkins B-cell lymphomas possess a deregulated bcl-2 gene resulting in a phenotype that is apparently resistant to programmed cell death (apoptosis). We have used a mouse lymphoma cell line (S49.1) that undergoes apoptosis in response to a variety of stimuli to determine the effect of bcl-2 expression on induction of apoptosis. S49 cells were stably transfected with recombinant amphotrophic retroviruses carrying either a G418 antibiotic resistance gene alone (S49-NEO) or this gene in combination with a bcl-2 complementary DNA (S49-Bcl-2). Three different agents previously shown to activate apoptosis by different pathways in S49 cells (dexamethasone, the calcium ionophore A23187, and cycloheximide) were used to examine the effect of bcl-2 expression on cell growth and apoptosis caused by multiple signal transduction pathways. Dexamethasone (DEX) treatment inhibited cell growth and stimulated cell death in S49-NEO cells. Although S49-Bcl-2 cells exhibited a similar antiproliferative response, they failed to die in response to steroid treatment. Western blot analysis revealed no difference in the levels of glucocorticoid receptor protein in the two cell lines, and both responded to glucocorticoid with a profound inhibition of protein synthesis. Cycloheximide (CX) and A23187 also had antiproliferative and cell killing effects in both cell types, although higher concentrations of each agent were needed to kill S49-Bcl-2 cells. To determine whether the loss of viability in response to these drugs was due to apoptosis, cells were examined morphologically and DNA integrity was examined by gel electrophoresis.(ABSTRACT TRUNCATED AT 250 WORDS)

Similar actions of glucocorticoids and calcium on the regulation of apoptosis in S49 cells.

Glucocorticoid-induced lymphocyte cell death is a programmed process which is thought to involve the calcium-dependent degradation of DNA into multiples of 180 basepairs, characteristic of internucleosomal degradation. We have used the glucocorticoid-sensitive mouse lymphoma cell line S49.1 [wild-type (wt)] and the glucocorticoid-resistant cell line S49.22r (nt-) to evaluate the role of both glucocorticoid receptors and calcium in the regulation of internucleosomal DNA degradation and expression of calcium-dependent deoxyribonuclease activity. DNA was isolated from untreated (control) and dexamethasone (dex)-treated viable cells and analyzed for internucleosomal DNA degradation by agarose gel electrophoresis, followed by ethidium bromide staining. Glucocorticoid treatment resulted in substantial internucleosomal DNA degradation in wt cells, but not in nt- cells. This effect was inhibited by coincubation of cells with dex and the glucocorticoid receptor antagonist RU486. In contrast to the glucocorticoid response, administration of either of two calcium ionophores, ionomycin or A23187, produced internucleosomal degradation of DNA in both wt and nt- cells, although the latter were less sensitive to ionophore treatment. Interestingly, A23187 treatment also resulted in a loss of cell viability in HeLa S3 cells, a cell line that does not exhibit glucocorticoid-induced apoptosis. No internucleosomal DNA degradation was detected in HeLa S3 cells killed by A23187. To determine whether similar nucleases are associated with this internucleosomal DNA degradation resulting from both glucocorticoid and calcium ionophore treatment, 0.3 M NaCl nuclear protein extracts were prepared from control and treated cells and analyzed for protein composition or nuclease activity. To assay for nuclease activity, nuclear extracts were electrophoresed in sodium dodecyl sulfate-polyacrylamide gels impregnated with [32P]DNA. Nuclease activity was detected by removal of sodium dodecyl sulfate from the gel, activation with calcium, and subsequent visualization of the loss of [32P]DNA by autoradiography. Dex treatment of wt cells resulted in the appearance of several proteins within the mol wt range of 12-18 kDa, only one of which (16-18 kDa) exhibited calcium-dependent nuclease activity. The appearance of these proteins in nuclear extracts was inhibited by coincubation of glucocorticoid-treated cells with RU 486. Glucocorticoid treatment did not result in the appearance of nuclease activity in nuclear extracts from nt- cells. Interestingly, A23187 or ionomycin treatment resulted in an increase in activity of the 16- to 18-kDa nuclease in both wt and nt- cells. These findings indicate that both glucocorticoid receptors and calcium may share common features in the regulation of apoptosis in lymphoid cells.

Changing factors and changing needs in women's health care.

The aforementioned social trends affecting women, including women in poverty, women in the labor force, and elderly women, are all ultimately related to problems of access to health care. In almost every age group, women use more health and medical services. Women are hospitalized more often, although their stays in hospitals tend to be shorter. Women also make more visits to health care providers for preventive health care, such as examinations and dental care. Access to care, however, is tied to ability to pay for the care. Medicaid payments for medical care are related to eligibility criteria in each state. Recent cuts in federal programs targeted eligibility for welfare and Medicaid. In 1982, 725,000 welfare recipients were declared ineligible. Given the earlier discussion of the predominance of women among those labeled poor in this country and the fact that two thirds of Medicaid recipients are women, these cutbacks have serious implications for women's health. Women are less likely to have medical insurance than men. Insurance coverage as a benefit is least likely to be offered in those areas where women work: part-time employment, small businesses, and manufacturing industries. Insurance eligibility is often dependent on a woman's marital status, despite the fact that 41.5 per cent of all American women are not spousal dependents. Insurance companies frequently adjust premiums for sex, age, income, race, and workforce characteristics, a policy which works against women. As the field of women's health expands and receives more emphasis, the data reflecting the experiences of large groups of women will have to be collected and analyzed ever more carefully. Information collected should include physiologic, psychosocial, and economic factors that together affect the health status of women. These data may then be used to guide health policy decision making, as well as provide a basis for health promotion and disease prevention interventions with individual clients.

PIP: As a result of decreases in maternal mortality and infectious diseases, women's life expectancy has increased rapidly in this century and is expected to reach 83 years by the year 2000. However, there are a large number of chronic conditions that negatively affect the quality of life of women today: urinary tract infection, menstrual cycle disorders, hypertension, diabetes, osteoporosis, arthritis, eating disorders, substance abuse, and mental depression. Although women's life expectancy is 7.5 years greater than that of men, the morbidity rates are significantly higher for women. As women continue to enter the labor force in large numbers, questions are being raised regarding the physical and psychological hazards of jobs traditionally considered to be women's work, the risks associated with jobs that are physically demanding or involve exposure to toxic substances, and the association between pregnancy outcome and employment. Further research is needed on the effects of multiple role stress on women's health. Another recent trend has been the feminization of poverty: 2/3 of all US adults classified as poor are women. The lack of financial resources has a detrimental effect on nutrition, access to health care, and other preventive behaviors. Yet another social change related to women's health is the increasing number of elderly in the population. Women comprise 72% of the elderly poor, and over 80% of all retiring female workers do not have pension benefits. Access to, availability of, and payment for health care are problems for elderly women. It is important that research address the physiologic, psychosocial, and economic factors that together affect women's health status.

The impact of adolescent females' assessments of parenthood and employment on plans for the future.

PIP: This study explores adolescent females' assessments of the rewards and costs of employment and parenthood, and the impact of these assessments on their plans for organizing their adult lives. 549 young women enrolled in the 11th and 12 grades at public schools in New York City completed questionnaires addressing their future work and fertility plans, characteristics of their present family life, and their assessments of how rewarding and costly the roles of employee and parent would be to them. Results suggest that daughters of homemakers and daughters of employed women differ in their assessment of each role, and take different factors into consideration when making these assessments. In evaluating work, daughters whose mothers are employed were likely to depend on their mothers' experiences for guidance. On the other hand, daughters whose mothers are homemakers have to go through a more elaborate process of deduction. They appear to assess whether their mothers are happy where they are now and then project whether another role would be more satisfying. These young women also tend to incorporate the opinion of the father more than do daughters of employed mothers. In evaluating the impact of children, the young woman's relation with her own mother is a critical factor affecting both group' assessments of the costs of children as well as the employed group's assessments of the rewards of children. Finally, the limitations of the sample should be kept in mind when discussing the results. Though the sample was large numerically, the overall response rate was low. In addition, the sample consisted of a large number of Catholic respondents.^ieng