From Creutzfeldt-Jakob disease to the mad cow epidemic.

Hans-Gerhard Creutzfeldt and Alfons Jakob independently authored clinical and pathologic descriptions of a new syndrome in the 1920s. This syndrome, which subsequently came to be named after them, was characterized by dementia, motor and coordination abnormalities, a fatal course, and pathologic findings of diffuse spongiform neuronal degeneration. Although it appeared for many years to be little more than a medical curiosity, Creutzfeldt-Jakob disease attained widespread attention by its pathologic similarity to kuru and bovine spongiform encephalopathy, "mad cow disease." Because there are sporadic, familial, and iatrogenic forms of Creutzfeldt-Jakob disease, it is considered to have both genetic and infectious aspects. Although its causation has for some time been ascribed to "slow viruses," the etiology of Creutzfeldt-Jakob disease is currently thought to be due to prions, small proteinaceous infectious particles that have genetic encoding. The debate regarding whether the appearance of atypical Creutzfeldt-Jakob disease can be linked to the epidemic of "mad cow disease" is currently unresolved.

Willy Burgdorfer: Lyme disease.

The documented history of erythema migrans dates to 1909, when Arvid Afzelius described a case of this skin lesion at a dermatologic meeting in Sweden. He felt that the eruption was likely produced by the bite of a tick. The initial description of Lyme arthritis appeared in 1977, and a number of the patients described in this series developed a rash thought to be erythema migrans. Four years later, Burgdorfer discovered the presence of spirochetes (subsequently named Borrelia burgdorferi) in ticks from an endemic locus of Lyme arthritis and determined this to be the causative organism of the disease. Lyme disease is the most common tickborne infection in the United States. Its natural course has been divided into three clinical stages. The infection begins with a rash and flulike symptoms and may progress after days to weeks to a disseminated stage and in months to years to a late stage. There is little information (except erythema migrans) about the clinical features of the illness that is specific for Lyme disease. There are a number of effective antibiotic treatment regimens, and although acute infection generally responds well to treatment, management of chronic illness with antibiotics has been less consistently successful. With respect to antibiotic prophylaxis, the few studies performed have led to the conclusion that, even in endemic areas, the risk of infection is so low that routinely instituting treatment following a tick bite is not warranted.

Transcriptional activation of the c-myc proto-oncogene in murine keratinocytes enhances the response to epidermal growth factor.

To investigate the relationship between activation of the c-myc proto-oncogene and the controls of cellular growth and differentiation of epidermal cells, a transcriptionally activated c-myc gene (DM-myc) was introduced into the established murine keratinocytes, BALB/MK. Exponential growth rates of myc-transfectants were not significantly different from that of parental BALB/MK cells. C-myc RNA transcripts were not detectable in confluent, mitogen-deprived cultures of parental BALB/MK cells, whereas four out of five clones expressed elevated levels of myc mRNA under these conditions. All of the cell lines, however, displayed density-dependent growth arrest in the G0/1 phase of the cell cycle. Maximal stimulation of quiescent BALB/MK cells with epidermal growth factor (EGF) caused a 70- to 100-fold increase of [methyl-3H]-thymidine incorporation into DNA. In the four subclones that expressed the myc gene, the peak thymidine incorporation into DNA was significantly higher than in BALB/MK cells, ranging from 340- to 650-fold control levels. This increased sensitivity to EGF was not due to autocrine mitogenic activity or to a change of EGF binding. Type beta transforming growth factor strongly inhibited the EGF-induced DNA synthesis in BALB/MK cultures as well as in each of the five transfectants (IC50 4-40 pM). Furthermore, both BALB/MK cells and the transfected subclones could be induced to form cornified cell envelopes by increasing the extracellular concentration of calcium. Thus, the constitutive expression of c-myc in BALB/MK appears to affect predominantly the reinitiation of DNA synthesis by EGF.

Down-regulation of glucocorticoid binding sites by retinoic acid in squamous carcinoma cells resistant to the induction of keratinization by hydrocortisone.

Physiologic concentrations of retinoic acid strongly inhibit the in vitro maturation of human squamous carcinoma cells in serum-free medium. Differentiation, as measured by the capacity to synthesize cornified cell envelopes, could be induced by hydrocortisone in retinoic acid-treated SqCC/Y1 and CE-81T cells. However, two other cell lines (C4-1 and A431) were less competent to spontaneously form cornified cell envelopes and resistant to the induction of envelope competence by hydrocortisone in the presence of retinoic acid. To investigate the mechanism underlying the resistance of these two lines to hydrocortisone, the characteristics of glucocorticoid receptors were analyzed. Whole cell dexamethasone binding sites ranged from 1300 to 9000 sites per cell for the four cell lines. The binding affinity for dexamethasone was similar in all four squamous carcinoma cell lines (1.32 to 4.75 nM). During retinoic acid-treatment, the binding of dexamethasone by intact SqCC/Y1 and CE-81T cells increased 1.5- to 3.0-fold over 48 h. In contrast, the number of dexamethasone binding sites were decreased by 80% in retinoic acid-treated A431 and C4-1 cells. In each case, the regulation of dexamethasone binding was dependent on the concentration of retinoic acid, with maximal effects being observed at 10(-6) M. Thus, the manner in which retinoic acid regulates the availability of dexamethasone binding sites might explain, in part, the effects of glucocorticoids on differentiation of retinoic acid-treated squamous carcinoma cell lines.

Reinitiation of DNA synthesis in quiescent mouse keratinocytes; regulation by polypeptide hormones, cholera toxin, dexamethasone, and retinoic acid.

Cloned mouse keratinocytes (MK-1 cells) display density-dependent growth arrest when reaching confluency in a serum-free medium with a calcium concentration less than 0.1 mM, supplemented only with insulin and transferrin. In this quiescent state, greater than 95% of the cell population is in the Go/1 phase of the cell cycle. Treatment of quiescent MK-1 cells with 1 to 10 ng/ml epidermal growth factor (EGF) resulted in a sharp burst of DNA synthetic activity. Both insulin and cholera toxin potentiated the mitogenic effect of EGF, but neither agent was necessary or sufficient to induce thymidine incorporation into DNA. Dexamethasone abolished the effect of insulin, but not the mitogenic effect of EGF alone. In contrast, retinoic acid (RA) did not possess any mitogenic effect for quiescent MK-1 cells, nor did it modulate the actions of EGF or dexamethasone. A number of commercially available crude extracts of bovine brain and pituitary were also capable of initiating DNA synthesis in resting MK-1 cells. Finally, transforming growth factor type beta (TGF beta) proved to be a potent inhibitor of the mitogen-induced DNA synthesis in MK-1 cells (IC50:10 pM). This defined culture system is eminently suited to study the regulation of DNA synthesis of epidermal cells. In addition, it can be used as a sensitive bioassay for the detection of epidermal mitogens, as well as inhibitors of DNA synthesis such as TGF beta.