TOP2A gene copy gain predicts response of epithelial ovarian cancers to pegylated liposomal doxorubicin: TOP2A as marker of response to PLD in ovarian cancer.

OBJECTIVE: The treatment of platinum resistant/refractory epithelial ovarian cancer (EOC) is a challenge for oncologists. One of the most utilized drugs in these patients is pegylated liposomal doxorubicin (PLD). As PLD is active only in a small subset of patients and causes side effects, selection of responsive patients is an unmet need and might be guided by the status of the DNA topoisomerase II alpha (TOP2A) that is poisoned by the drug. METHODS: From 176 ovarian cancers treated in three institutions, we selected 38 patients treated with PLD monotherapy as second/third line of treatment. TOP2A gene copies were measured using Fluorescent In Situ Hybridization (FISH) and expression evaluated using immunohistochemistry. Patients' derived xenografts (PDXs) of ovarian cancers were used to assess the correlation between TOP2A protein expression and response to PLD. RESULTS: Clinical data showed that TOP2A gene gain that is paralleled by increased expression of the protein, was associated with a higher probability of clinical benefit from PLD. Treatment of PDXs demonstrated that only xenografts showing a high percentage of TOP2A expressing cells underwent tumor shrinkage when treated with PLD. CONCLUSIONS: These data show that TOP2A gene gain and protein over-expression might predict activity of PLD in platinum resistant/refractory EOC.

Apoptosis of L929 cells by etoposide: a quantitative and kinetic approach.

Exponentially growing L929 cells were continuously exposed to 1 or 10 microM etoposide (VP-16). The effects of such treatment on cell growth, cycle distribution, morphology, and selected biochemical events were examined. DNA synthesis rates were markedly decreased and the protein/DNA ratio increased (unbalanced growth). Growth was blocked, with most cells being cycle arrested by 24 h in (late S-)G2-M. An asynchronous process of cell death then developed. Cells initially shrank into eosinophilic, trypan blue-excluding bodies, which were then released into the medium, and eventually became permeable to trypan blue. Transmission electron microscopy confirmed that dying cells acquired an apoptotic morphotype, with compaction and margination of chromatin, loss of microvilli, and shrinkage of cytoplasm and nucleus. Tissue transglutaminase activity and intensity of immunostaining rapidly increased in treated cultures. Internucleosomal DNA fragmentation could not be detected by agarose gel electrophoresis, yet flow cytometry revealed that the apoptotic bodies had a very low DNA fluorescence (< or = 10% of the 2n value). In agreement with the microscopic findings, this suggested that extensive DNA degradation had occurred in dead cells. While rates of cell loss from the monolayer amounted to 21 and 57% day(-1) (1 and 10 microM VP-16, respectively), apoptotic indexes largely underestimated the extent of the process. These indexes only measured the accumulation of apoptotic bodies, i.e., the balance between their generation and disposal. The latter occurred by mechanisms similar to those that operate in tissues: "secondary necrosis" or phagocytosis by viable homotypic cells in the monolayer ("homophagy").

Presynaptic co-localization of carnosine and glutamate in olfactory neurones.

Olfaction plays a dominant role in modulating behaviour in most vertebrate species and the olfactory bulb is considered a model system for characterizing principles of neural computation. Nevertheless, although the physiology and neurochemistry of the olfactory circuits have been widely studied, the neurotransmitter released by olfactory receptor neurones remains unknown. We now describe the ultrastructural localization of the dipeptide carnosine and the excitatory amino acid glutamate in the glomerular layer of the mouse olfactory bulb. We demonstrate that both carnosine-like and glutamate-like immunoreactivities are selectively co-localized in the olfactory neurone boutons. These observations, taken with the recent findings of glutamate-receptor subunit expression in rodent olfactory bulb, argue compellingly for a role of glutamate in olfactory neurotransmission and suggest a modulatory effect of carnosine.

Double labeling with non-isotopic in situ hybridization and BrdU immunohistochemistry: calmodulin (CaM) mRNA expression in post-mitotic neurons of the olfactory system.

We describe a novel histochemical procedure for simultaneous detection of mRNA expression by in situ hybridization (ISH) and DNA synthesis on cells that are pulse-labeled with bromodeoxyuridine (BrdU) by immunohistochemistry (ICC). Pregnant rats were injected with BrdU at embryonic Day 20 and the olfactory bulbs of their pups were collected daily. The expression of calmodulin (CaM) mRNA was analyzed by ISH with an anti-sense digoxigenin-labeled riboprobe and BrdU incorporation by indirect ICC. Starting 5 days after BrdU injection, a few tufted and granular neurons of the olfactory bulb were observed to be double labeled for CaM mRNA and BrdU. To study the olfactory neuroepithelium, adult animals were injected with BrdU, sacrificed after 30 days, and the nasal mucosa dissected and decalcified. The co-expression of CaM mRNA and BrdU incorporation was then analyzed in the olfactory neuroepithelium: BrdU-positive primary olfactory neurons were also CaM mRNA positive. The combination of ISH and ICC on the same section resulted in improved BrdU staining with respect to both increased intensity and reduced background levels. The procedure described here can be applied to a variety of problems in developmental biology and is of potential value for correlating the timing of specific mRNA expression with the birth date of a cell type of interest.

[Effect of street noise on blood catecholamines, cyclic AMP and various cardiovascular and metabolic functions in a group of subjects with untreated essential hypertension]. / Effetti del rumore stradale sulle catecolamine ematiche, sul cAMP e su alcune funzioni cardio-vascolari e metaboliche in un gruppo di soggetti ipertesi essenziali non trattati.

The effect of road noise on blood catecholamines, cAMP and certain cardiovascular and metabolic parameters was studied in a group of young untreated essential hypertensives. It was found that the 10' stimulus increased both systolic and diastolic pressure values. Blood catecholamines rose only after 5'. The most significant increase was in adrenaline as opposed to noradrenaline and dopamine. Significant changes were noted in cAMP and triacylglycerols (10' and 15' after commencement of the stimulus respectively). Comparison with previous results in normotensives suggested that the catecholamine response to stress is primarily alpha-receptorial when blood pressure is normal and beta-adrenergic in hypertension.

[Myxoma of the left atrium. General considerations and description of a case]. / Mixoma dell'atrio sinistro. Considerazioni generali e descrizione di un caso.

A case of left atrial myxoma is described. The neoplasia was observed in a female with no previous rheumatic history in whom cardiopathy was diagnosed only a few months before hospitalisation. The rapid and erratic course of the cardiopathy, the fact that changes of posture modified objective symptoms, the presence of dyspnoea and peripheral microembolisms were all clinical indicators of a possible myxoma, later confirmed by echography and cardiac catheterism. The anatomical and functional characteristics of myxomas are also discussed.

[Effects of street noise on plasma catecholamines, cyclic AMP and various cardiovascular and metabolic functions in a group of normal subjects]. / Effetti del rumore stradale sulle catecolamine ematiche, sul cAMP e su alcune funzioni cardio-vascolari e metaboliche in un gruppo di soggetti normali.

Exposure to traffic noise for ten minutes increased diastolic pressure without affecting systolic pressure and heart rate. Plasma catecholamine levels were enhanced only five minutes after noise stimulation. Noradrenaline increased more significantly than dopamine and adrenaline. No changes in plasma cAMP levels and hematochemical parameters ( cholesterol , glucose, triglycerides , uric acids) were observed. Data suggest that traffic noise can be considered a stress-full stimulus and its effect on diastolic pressure can be correlated to alpha-receptor stimulation.

[Rupture of the chordae tendineae and mitral prolapse. General considerations and description of a case]. / Rottura di corda tendinea e prolasso mitralico. Considerazioni generali e descrizione di un caso.

A severe case of acute mitral insufficiency caused by a ruptured tendinous cord is described. The subject had an unrecognised mitral prolapse and had been suffering from tachycardia for years when an attack of pulmonary oedema occurred some days after apparently well tolerated exertion. The phenomena behind the event are discussed with particular reference to the mechanisms, closure and distribution of prolapses in so-called mixamatosic degeneration of the valve. The importance of body position and exertion in the creation and deterioration of prolapses is also emphasised.