Microcalcifications in breast cancer: novel insights into the molecular mechanism and functional consequence of mammary mineralisation.

BACKGROUND: Mammographic microcalcifications represent one of the most reliable features of nonpalpable breast cancer yet remain largely unexplored and poorly understood. METHODS: We report a novel model to investigate the in vitro mineralisation potential of a panel of mammary cell lines. Primary mammary tumours were produced by implanting tumourigenic cells into the mammary fat pads of female BALB/c mice. RESULTS: Hydroxyapatite (HA) was deposited only by the tumourigenic cell lines, indicating mineralisation potential may be associated with cell phenotype in this in vitro model. We propose a mechanism for mammary mineralisation, which suggests that the balance between enhancers and inhibitors of physiological mineralisation are disrupted. Inhibition of alkaline phosphatase and phosphate transport prevented mineralisation, demonstrating that mineralisation is an active cell-mediated process. Hydroxyapatite was found to enhance in vitro tumour cell migration, while calcium oxalate had no effect, highlighting potential consequences of calcium deposition. In addition, HA was also deposited in primary mammary tumours produced by implanting the tumourigenic cells into the mammary fat pads of female BALB/c mice. CONCLUSION: This work indicates that formation of mammary HA is a cell-specific regulated process, which creates an osteomimetic niche potentially enhancing breast tumour progression. Our findings point to the cells mineralisation potential and the microenvironment regulating it, as a significant feature of breast tumour development.

Evidence that the acute behavioral and electrophysiological effects of bupropion (Wellbutrin) are mediated by a noradrenergic mechanism.

Bupropion (BW 323U66) has been considered a dopaminergic antidepressant based on its ability to inhibit the uptake of dopamine (DA) somewhat more selectively than it inhibits uptake of norepinephrine (NE) or serotonin (5-HT). This report describes new evidence that bupropion selectively inhibits firing rates of NE cells in the locus coeruleus (LC) at doses significantly lower than those that inhibit activity of midbrain DA cells or dorsal raphe 5-HT cells. The IC50 dose (13 mg/kg i.p.) for inhibition of LC firing produced plasma concentrations that were not significantly different from those generated by the ED50 in the Porsolt test (10 mg/kg i.p.). The fourfold higher dose needed to inhibit DA cell firing (IC50 = 42 mg/kg i.p.) was similar to the dose associated with locomotor stimulation in freely moving rats. Bupropion did not change the firing rates of 5-HT cells in the dorsal raphe nucleus at any dose. In both in vitro and in vivo tests, the metabolite 306U73 (hydroxybupropion), a weak inhibitor of NE uptake, was approximately equipotent to bupropion with regard to inhibition of LC cells. Another metabolite, 494U73, had no effect on LC firing rates over a wide range of doses. Because of species variation in metabolism, 306U73 was not detected in plasma of rats after i.v. doses of bupropion that inhibited LC firing. Only trace amounts of 306U73 were detected after bupropion dosing for the Porsolt test. Pretreatment with reserpine markedly depleted catecholamines and reduced (by 30-fold) the potency of bupropion to inhibit LC firing.(ABSTRACT TRUNCATED AT 250 WORDS)

Managing skin damage induced by doxorubicin hydrochloride and daunorubicin hydrochloride.

The pathophysiology and mechanisms of toxicity of anthracycline-induced skin damage are reviewed, and the various available therapeutic interventions are discussed. Skin ulcers caused by the vesicant antineoplastic agents doxorubicin hydrochloride and daunorubicin hydrochloride begin slowly, and the extent of the tissue damage produced is often underestimated. Within a week, untreated infiltrations of these agents can advance to serious indurations and ulcerations, causing extensive damage to underlying structures such as tendons and bones. Two theories have been proposed to explain the mechanism of action of anthrocycline-induced tissue damage; one holds that doxorubicin-DNA complexes form causing cell death, and the other holds that these agents are reduced to free radicals that can cause cell-membrane damage. Nonpharmacologic treatment of extravasation consists of stopping the infusion at the first sign of a problem and attempting to aspirate fluid and drug back through the same needle. The application of ice packs for the next 24-72 hours is recommended. A variety of pharmacologic approaches have been evaluated to ameliorate tissue damage. Corticosteroids, sodium bicarbonate, beta-adrenergic agents, and dimethyl sulfoxide have been used with some success. Patients who do not respond to initial conservative treatments should be referred to a plastic surgeon for skin grafting and reconstruction. The best treatment for anthracycline toxicity is prevention.

Effects of estrogen and progesterone on transcription, chromatin and ovalbumin gene expression in the chick oviduct.

The effects of estrogen, progesterone and estrogen + progesterone combined on nuclear transcriptional processes in oviducts of immature chicks, previously withdrawn from estrogen, are reported. The responses to the steroids of the endogenous nuclear RNA polymerase activities, both nucleolar (I) and nucleoplasmic (II), the chromatin compositions and template capacities, and the appearance of ovalbumin messenger RNA (mRNA) are compared. When immature chicks (previously treated at 14 days with estrogen) are withdrawn from estrogen treatment, there is a gradual reduction in both polymerase activities. Diurnal variations in polymerase II activties in the oviduct of withdrawn chicks required that subsequent experiments include time-matched controls. The hormones alter RNA polymerase II and II activities in vivo as assayed in isolated nuclei. Progesterone represses the polymerase I and II activities, while estrogen alone and estrogen + progesterone enhance both polymerase activities immediately after injection. Diethylstillbestrol, a synthetic estrogen, causes changes similar to those of estrogen. The effects of these steroids on the polymerases are detected within 15 min of hormone injection. Changes in the capacities of chromatins to serve as template for RNA synthesis in general correlated with changes in polymerase II activities. Interestingly, in the case of estrogen treatment, the acidic chromatin protein (but not histone) levels fluctuate positively with the template capacities of the chromatin. An antagonism between estrogen and progesterone is observed in the responses of both RNA polymerases I and II activities as well as in the chromatin template capacity. Levels of messenger RNA coding for ovalbumin, as detected by hybridization with labeled complementary DNA, increase in oviducts of withdrawn chicks within 2--3 of the injection of estrogen, progesterone or estrogen + progesterone. This rapid accumulation of ovalbumin mRNA is not accompanied in each case by a similar increase in polymerase II activity or chromatin template capacity.

Quantitation of elongating form A and B RNA polymerases in chick oviduct nuclei and effects of estradiol.

The number of elongating form A and B RNA polymerases in chick oviduct nuclei was estimated by measuring incorportaion of 3H-UTP into 3' termini of nascent RNA chains in the presence of heparin to inhibit initiation, and quantitation labeled uridine released from these termini after alkaline hydrolysis. The method corrects for conversion of UMP to uridine (U) during manipulations and for production of 3' termini by ribonucleases and phosphatases. The results indicate that a large fraction of RNA polymerases elongating in vivo is retained in isolated nuclei: per diploid genome, approximately 1 x104 form B and 2 x103 form A enzymes are present. These levels are sufficient to maintain normal in vivo rates of mRNA and rRNA synthesis, but the average density of packing of polymerases on DNA is considerably less than the maximum density predicted by Miller and Bakken (1972), suggesting that initiation of polymerases of DNA is a limiting factor in the control of transcription. Rates of elongation of polymerases in vitro are severely impaired, indicative of a loss of elongation factors during nuclear isolation. After 6 hr of estradiol treatment in vivo, the level of form A enzymes in ovoduct nuclei increases to 5 x 103; little change in the number of form B enzymes is seen.

Effects of progesterone on protein metabolism in chicken oviduct tissue pretreated with oestrogen.

1. The effect of simultaneous injections of oestrogen benzoate and progesterone (0.5mg/day each) on immature chicken oviduct tissue pretreated with oestrogen benzoate (0.5mg/day) was studied. 2. After 3 days of treatment with both hormones, the weight of the tissue doubles as compared with tissue treated only with oestradiol benzoate. 3. The progesterone-induced weight increase had no effect on total DNA content of the tissue, but greatly increased the protein/DNA and RNA/DNA ratios. 4. Amino acid incorporation in vivo after progesterone treatment was elevated as measured by using 2h pulses of amino acids; this effect could be accounted for by observed alterations in the concentrations of free amino acids in the tissue. 5. With longer pulses of amino acid the specific radioactivity of total protein remained high in tissue treated with progesterone, at times when specific radioactivity of protein in oestrogen-treated animals was diminishing. 6. From a knowledge of the specific radioactivity of labelled amino acids in the free amino acid pool and in newly synthesized protein, the rate of protein synthesis was estimated in differently treated tissues. 7. The results suggest that progesterone treatment does not cause an increase in protein synthesis. It is concluded that the observed accumulation of oviduct protein is achieved via an effect of progesterone on protein catabolism.