The strategic role of health informatics in integrated delivery systems.

Having accurate measures and high-quality health information is critically important for all providers today. Integrated delivery systems are faced with increasing demands for numerous redundant, sometimes conflicting, performance measurement and reporting data from managed care customers, regulators, and accreditors. When implemented independently within each organizational subunit, these measurement systems are costly and difficult to manage. Centralization of all measurement services can maximize the productivity of the costly resources required to deliver them and can achieve efficiencies, cost savings, and a better balance between internal and external resources while collecting information that is of a higher quality for managerial and clinical decision making.

The cellular oncogene p53 can be activated by mutagenesis.

P53 is a cellular phosphoprotein of short half-life (t1/2) which is present at elevated levels in cells transformed by various stimuli including viruses, chemicals and radiation. p53 forms specific stable complexes with simian virus 40 (SV40) large-T antigen and an adenovirus E1b protein of relative molecular mass (Mr) 57,000. A number of reports have associated p53 with cell proliferation, and p53 complementary DNA expression constructs immortalize primary cells in vitro and render them sensitive to transformation by an activated ras oncogene. We have examined the biological properties of a set of p53 expression constructs, and report here that cellular immortalization by a wild-type p53 cDNA gene is conditional upon the promoter/enhancer construction used, but that p53 can extend cellular lifespan by a second distinct mechanism involving rearrangements of the coding sequence which give rise to stable protein products. Cells immortalized by one of these mutants are refractory to subsequent transformation by a ras oncogene, indicating that cellular immortalization and ras cooperation are separate activities.

Activation of N-ras in a human melanoma cell line.

DNA isolated from cell line Mel Swift, a human melanoma cell line, transforms NIH3T3 cells. Southern blot analysis of DNA from secondary foci revealed conserved 8.8- and 7.8-kilobase EcoRI fragments which hybridized with a human repetitive sequence clone, blur 8. The activated transforming gene was identified as N-ras, and the 8.8-kilobase EcoRI fragment from a secondary transformant was cloned. Synthetic 17-mer oligonucleotides which spanned either the normal codon 61 (CAA) or a mutant codon 61 (AAA) were used for hybridization. Cloned N-ras from melanoma cell line Mel Swift hybridized to the mutant (AAA) oligonucleotide. From this we predicted a glutamine-to-lysine substitution in amino acid 61, a change confirmed by conventional sequencing of the first and second exons of N-ras from cell line Mel Swift. Transfection experiments showed that only those recombinant clones with the mutation in position 61 were biologically active.

Cellular immortalization by a cDNA clone encoding the transformation-associated phosphoprotein p53.

Malignant transformation of primary cells requires at least two distinct and characteristic alterations in cellular behaviour. The first, cellular immortality, can be induced by chemical carcinogens or by cloned oncogenes such as polyoma large T (ref. 4), adenovirus early region 1A (E1A) or the oncogene from avian (MC29) myelocytomatosis virus, v-myc. Cells whose in vitro life-span has been extended by these procedures can be fully transformed by transfection with oncogenes belonging to a different complementation group, including genes of the ras family, adenovirus E1b and polyoma virus middle T (refs 4, 5). The unstable cellular phosphoprotein p53 is frequently present at elevated levels in transformed cells and is stabilized by the formation of complexes with simian virus 40 (SV40) large T or adenovirus E1b 57K protein. Although several reports have associated p53 with cell proliferation, its role remains obscure. We have cloned complementary DNA sequences encoding murine p53 and report here that transfection of p53 expression constructs into cells of finite lifespan in vitro results in cellular immortality and susceptibility to transformation by a ras oncogene.

A novel transforming gene in a human malignant melanoma cell line.

Cellular transforming genes can be detected in human tumours by DNA-mediated transfection into NIH 3T3 mouse fibroblasts. The activated transforming genes have been, in most cases, members of the ras gene family, of which the most frequently found is the c-Ki-ras oncogene and least frequently the c-Ha-ras gene. An increasing number of studies has identified the presence of activated N-ras (which has no known viral homologue) in human tumour cell lines. Furthermore, other transforming genes, distinct from the ras gene family, have been reported in B-and T-cell lymphomas. The activation of c-Ha-ras and N-ras has been described in some cell lines derived from cases of human malignant melanoma. Here we describe the presence of transforming activity in the DNA from a human melanoma cell line which shows weak homology with members of the ras oncogene family.

Promotion of fibrosarcoma cell growth by products of syngeneic host macrophages.

Cells from a C57BL/cbi chemically induced fibrosarcoma (FS6) require exogenous platelet-derived growth factor (PDGF) for in vitro proliferation (as do normal "untransformed" fibroblasts) whereas cells obtained from the FS6M1 tumour, a spontaneous metastasizing subline, show autonomy from PDGF in vitro. Furthermore, the FS6 cells exhibit very low colony formation in an anchorage-independent growth assay. In vivo, this tumour is immunogenic, rarely metastasizes and is heavily infiltrated by host macrophages. Studies of in vitro cell proliferation and anchorage-independent growth show that syngeneic host macrophages from the peritoneal cavity or from the growing tumour release a diffusible factor(s) which has (1) growth-stimulating activity on FS6 cells in monolayer cultures in PDGF-poor medium and (2) potent colony-stimulating activity on FS6 cell cultured in methyl-cellulose-containing medium. These macrophage supernatants stimulate proliferation of quiescent normal fibroblasts in monolayer culture as well as FS6 sarcoma cells, but do not stimulate anchorage-independent growth of normal cells. Supernatants from BCG-elicited macrophages were shown to contain abundant arginase, and were cytolytic to FS6 cells but not to normal cells. Heat inactivation abrogated the arginase and cytotoxicity, revealing heat-stable mitogenicity for FS6 cells and normal fibroblasts. The stimulatory effect of macrophages on FS6 sarcoma cells can be mimicked by the addition of the tumour promoter 12-tetradecanoyl-phorbol-13-acetate (TPA) and supports the hypothesis that macrophages could play a significant role in multistage carcinogenesis by providing a source of endogenous promoter.

Platelet-derived growth-factor requirements for in vitro proliferation of normal and malignant mesenchymal cells.

Serum obtained by clotting whole blood contains a potent mitogen with apparent specificity for mesenchymal cells. This peptide wound-healing hormone, derived from platelets, is known as platelet-derived growth factor (PDGF). Serum obtained by clotting plasma contains no detectable growth-promoting activity for fibroblasts, and is therefore a valuable additive to culture medium for an examination of the autonomy of cells from exogenous PDGF. Fibroblasts from man, mouse and hamster remain mitotically quiescent in plasma-derived serum and proliferate only when a source of PDGF is added. Normal human kidney epithelial cells and human T-cells proliferate normally in plasma-derived serum, and are unaffected by the addition of PDGF. A range of virally transformed cells and malignant cells from chemically induced rodent sarcomas was tested for their proliferative capacity in plasma-derived serum and their response to exogenous PDGF. A complete spectrum of PDGF-dependence was revealed. Polyoma-transformed BHK21 cells and SV40-transformed 3T3 cells showed complete PDGF independence. Cells from 7 chemically induced rat or mouse sarcomas provided results which ranged from the FS6 (a C57BL Cbi mouse sarcoma which was completely PDGF dependent) to MC28 (a hooded rat sarcoma) which was completely PDGF independent. The dependence of proliferation of these cells on PDGF showed a close correlation with several features of their in vivo behaviour. Tumours which were non-immunogenic in syngeneic hosts, contained few host macrophages and produced a high incidence of spontaneous distant metastases provided PDGF-independent cells. Cells from highly immunogenic, macrophage-rich "non-metastasizing" tumours were on the other hand PDGF dependent and tumours of intermediate "malignancy" provided cells with partial autonomy from PDGF. An assay for anchorage-independent growth provided data which also correlated with autonomy from PDGF. However, daily addition of large amounts of PDGF to BHK21 C13 cells induced reversible anchorage independent growth. The value of plasma-derived serum for the investigation of the proliferative autonomy of malignant cells is emphasized.

Immunity as the predominant factor determining metastasis by murine lymphomas.

The metastatic behaviour of the L5178E (non-M) lymphoma and a highly metastatic subline L51787ES (M) were studied in syngeneic DBA2 mice. The non-M tumour rarely metastasizes in intact syngeneic mice, but produces extensive and rapidly lethal metastases when implanted into irradiated recipients. The metastatic behaviour of the M subline is unaffected by irradiation of the host. By conventional transplantation criteria, the non-M tumour is more immunogenic than the M subline. Both tumours, however, produce similar responses in a lymphnode weight-gain assay. Host-cell infiltration of the tumours growing s.c. is much greater in the non-M than the M, the infiltrating cells being Fc-receptor-positive and maturing into macrophages after 2 days in vitro. Although spontaneous in vitro motility of the M cells is much greater than that of the non-M, the metastatic behaviour of the tumours is clearly determined by host immunological responses.