Apoptosis and growth arrest induced by platinum compounds in U2-OS cells reflect a specific DNA damage recognition associated with a different p53-mediated response.

Mononuclear and multinuclear platinum complexes are known to induce distinct types of DNA lesions and exhibit different profiles of antitumor activity, in relation to p53 mutational status. In this study, we investigated the cellular effects of exposure to two platinum compounds (cisplatin and the multinuclear platinum complex BBR 3464), in the osteosarcoma cell line, U2-OS, carrying the wild-type p53 gene and capable of undergoing apoptosis or cell cycle arrest in response to diverse genotoxic stresses. In spite of the ability of both compounds to up-regulate p53 at cytotoxic concentrations, exposure to BBR 3464 resulted in cell cycle arrest but only cisplatin was capable of inducing significant levels of apoptosis and phosphorylation at the Ser15 residue of p53. The cisplatin-induced protein phosphorylation, not detectable in cells treated with BBR 3464, was associated with RPA phosphorylation, a specific up-regulation of Bax and down-regulation of p21(WAF1). Cells treated with BBR 3464 displayed a different cellular response with evidence of cytostasis associated with a high induction of p21(WAF1). The regulation of p21(WAF1) after cisplatin or BBR 3464 exposure required a p53 signal, as documented using stable transfectants expressing a dominant-negative form of p53 (175(his)). Taken together, these results indicate that cellular response to different genotoxic lesions (i.e. apoptosis or growth arrest) is associated with a specific recognition of DNA damage and a different p53-mediated signaling pathway. Multinuclear platinum complexes could be regarded as useful tools for investigating the p53-mediated process of cell cycle arrest in response to DNA damage.

Hemoglobin near-infrared spectroscopy and surface EMG study in muscle ischaemia and fatiguing isometric contraction.

BACKGROUND: To assess how muscle ischaemia and isometric fatiguing contraction influence oxygen content in striated muscle. METHODS: We simultaneously measured changes in hemoglobin near-infrared (NIR) spectroscopy and in surface EMG before, during, and after muscle ischaemia and ischaemia plus muscle isometric fatiguing contraction. Seventeen healthy male subjects (age range: 19-40 yrs) were examined in our Clinical Neurophysiology Unit. Test I (9 subjects): hemoglobin NIR spectroscopy and stimulated surface EMG were measured for 2 minutes at rest, for 4 minutes during complete ischaemia of tibialis anterior muscle, and for twelve minutes during recovery. Test II (all subjects): hemoglobin NIR spectroscopy and surface EMG were measured for 2 minutes with the subjects performing brief non-fatiguing contractions, for 4 minutes with the subject performing maximal isometric contraction in complete ischaemia, and for twelve minutes during recovery. EMG parameters measured: median density frequency (MDF); muscle fiber conduction velocity (MFCV). NIR spectroscopy parameters measured: percentage of amplitude decrement (% AD) and nadir time (NT) during ischaemia and ischaemic effort; half-recovery time (1/2 RT) from ischaemia effort. RESULTS: At EMG, we observed a significant shift towards lower values of both MFCV and MDF during fatiguing isometric contraction. MDF recovery was faster then MFCV recovery. At NIR spectroscopy, the 1/2 RT slowed a fast pattern in twelve subjects and a slow pattern in five. A significant relationship was found between AD% and 1/2 RT values of test I and AD% and 1/2 values of test II. We found a positive relationship between NT and 1/2 RT in test II. CONCLUSIONS: Surface EMG and hemoglobin NIR spectroscopy can be applied simultaneously to evaluate both fatigue intensity and blood flow changes in striated muscle.

A novel 9-aza-anthrapyrazole effective against human prostatic carcinoma xenografts.

OBJECTIVES: Systematic investigation of a novel series of intercalating agents, 9-aza-anthrapyrazoles, has led to the identification of a promising analogue, BBR 3438. This study describes the antitumour efficacy of the novel compound in human prostate carcinoma models and the molecular/cellular basis of its activity. METHODS AND RESULTS: The novel 9-aza-anthrapyrazole BBR 3438 was significantly more effective than doxorubicin and losoxantrone (DuP-941) in two of the three tested prostate carcinoma models. The superior activity was more evident in PC3 tumour, since BBR 3438 produced an appreciable rate of complete tumour regressions. Under these conditions, the drug-induced antiproliferative activity paralleled delayed apoptosis. Tumour response to in vivo drug treatment was associated with an early down-regulation of Bcl-2, which was somewhat more marked for the aza compound. In fact, the 9-aza-anthrapyrazole induced DNA cleavage in vitro with isolated DNA topoisomerase II (isoform alpha) and DNA strand breaks in prostatic carcinoma cells. Although the molecular effects of losoxantrone and the 9-aza analogue on the enzyme target were comparable, the cytotoxic effects of BBR 3438 could be enhanced by long-term exposure as a consequence of favourable cellular accumulation and prominent DNA-binding affinity. In addition, a lower reduction potential of the 9-aza-anthrapyrazole in comparison with classical anthrapyrazoles suggests an increased ability of the drug to induce oxidative stress following free radical production, which may be a contributing factor in determining the long-term response (i.e. delayed cell death) to genotoxic damage. CONCLUSIONS: BBR 3438 exhibited a unique profile of preclinical activity with a superior efficacy against prostatic carcinoma models compared to reference compounds (doxorubicin and losoxantrone). The antitumour efficacy of BBR 3438 against prostatic carcinoma could be the result of a combination of favourable events, including enhanced intracellular accumulation and an increased DNA-binding affinity favouring the accumulation of multiple sublethal or lethal damage. In spite of its enhanced cytotoxic potency, the 9-aza compound was better tolerated in vivo than losoxantrone, thus improving the therapeutic index. The preclinical profile of efficacy against prostatic carcinoma, a tumour resistant to conventional antitumour drugs, makes the novel 9-aza-anthrapyrazole BBR 3438 a promising candidate for clinical evaluation.

Embutidos cocidos funcionales / Functional cooked sausage-type product

Se ha optimizado, desde el punto de vista funcional, un embutido cocido, útil para prevenir la aterosclerosis, usando aceites vegetales con alto contenido en ácidos grasos poliinsaturados.El producto obtenido a nivel artesanal y posteriormente a nivel industrial con el apoyo de importante frigorífico, no posee grasas animales adicionadas, no contiene ácidos grasos trans, tiene bajo contenido en sodio, es bajo en calorías, no presenta cambios organolépticos considerables con relación a productos similares de primeras marcas, y esta enriquecido con AGPIw3 obteniéndose una relación w6:w3 ideal.(AU)

Embutidos cocidos funcionales / Functional cooked sausage-type product

Se ha optimizado, desde el punto de vista funcional, un embutido cocido, útil para prevenir la aterosclerosis, usando aceites vegetales con alto contenido en ácidos grasos poliinsaturados.El producto obtenido a nivel artesanal y posteriormente a nivel industrial con el apoyo de importante frigorífico, no posee grasas animales adicionadas, no contiene ácidos grasos trans, tiene bajo contenido en sodio, es bajo en calorías, no presenta cambios organolépticos considerables con relación a productos similares de primeras marcas, y esta enriquecido con AGPIw3 obteniéndose una relación w6:w3 ideal.