Synchronized onset of nuclear and cell surface modifications in U937 cells during apoptosis

In this study we investigated the relationship between nuclear and cell surface modifications (i.e. blebbing, phosphatidylserine [PS] and sugar residues exposure) in a monocytic cell line, U937, during apoptosis induced by oxidative stress (1mM H2O2) or inhibition of protein synthesis (10 mg/ml puromycin). Dying cells were simultaneously observed for nuclear modifications, presence of superficial blebs and plasma membrane alterations. Morphological analysis performed by conventional fluorescence microscopy, or by transmission and scanning electron microscopy showed that the courses of nuclear and membrane alterations occured concomitantly, but the phenotype was dependent on the stage of the apoptotic process and the type of apoptogenic inducer used. The progression of apoptosis in U937 cells beyond early stages resulted in the extensive formation of blebs which concomitantly lost some typical markers of apoptosis, such as PS and sugar residues. Therefore, the modality by which the nucleus condenses, or the amount and the pattern of distribution of PS on the cell surface were, for each cell line, strictly related to the apoptogenic inducer. The morphological data reported in the present paper should lead to a more precise quantification of apoptosis by improving the detection of apoptotic cells in vivo (i.e. in tissue, organs), which is a crucial point in the evaluation of efficiency of antiproliferative drugs, such as antiblastic or immunosuppressive compounds

Performance of Delta4 Phantom+ using Flattening-Filter and Flattening Filter-Free beams

EnThe goal of this study was to evaluate the performances of the new pre-treatment system Delta4 Phantom+ in terms of a stable and sensitive tool for achieving a reliable verification. Delta4+ was evaluated for verification with 6FFFMV beams using static beam arrangement. The accelerator output was monitored with the ionization chamber. The device was tested for dose-rate dependence, linearity and stability using its daily output correction. The response of Delta4+ was evaluated for energy 6FFFMV, measuring the gamma index of four Volumetric Modulated Arc Therapy (VMAT) plan. The response of Delta4+ as a function of accelerator dose rate is in agreement with the ionization chamber with a difference smaller than 0,1%. The output is constant for different MU. VMAT plan analysis show values within 98,3%-100% with a threshold of 3%-3mm, while with a threshold of 2%-2mm the values are within 93,5%-97,5%. Delta4+ is an accurate device. For all the measurements made, uncertainties below 1% were obtained.ItLo scopo di questo studio è stato quello di valutare la risposta in termini di stabilità e sensibilità del nuovo sistema pre-trattamento Delta4 Phantom+. La risposta del sistema è stato studiata utilizzando fasci di fotoni da 6MVsenza filtro di flattening (6FFFMV). L'output dell'acceleratore è stato monitorato con una camera a ionizzazione. La dipendenza dal dose rate, la linearità e la stabilità sono state verificate utilizzando il fattore di correzione giornaliera fornito dal sistema. La risposta è stata verificata su 4 piani di trattamento con tecnica VMAT. La risposta del Delta4+ al variare del dose rate è in accordo con quella della camera a ionizzazione con una scarto minore dello 0.1%. La risposta al variare delle UM è costante con una differenza entro lo 0.4%. L'analisi dei piani VMAT mostra valori di indice gamma per soglia 3%-3mm compresi tra 98.3%-100%, mentre per 2%-2mm i valori sono tra 93,5%-97,5%. Il Delta4+ risulta essere un sistema accurato in quanto l'incertezza ottenuta su tutte le misure non supera l'1%