Advances in the treatment of pancreatic neuroendocrine tumours.

Pancreatic neuroendocrine tumours (pNETs) are relatively rare and generally felt to follow an indolent course. But poorly differentiated tumours can behave aggressively with 5-year survival ranging from 31% to 48%. Recent data suggest that patients with pNETs may derive benefit from treatment targeting the molecular changes expressed in this tumour group. This article describes advances in the treatment of unresectable pNETs that have led to a doubling of progression free survival.

Diversity of peripheral blood mononuclear cells as revealed by a novel multiple microgel "comet assay".

Multiple microgel comet assay (MMCA) is a metho-dological adaptation of the single-cell gel electrophoresis assay in which we have introduced the use of standard agarose plug molds in an attempt to improve and expand the applications of the assay. We focused on the study of the heterogeneity of peripheral blood mononuclear cells (PBMC) at the level of the basal single-strand breakage and the DNA damage induction caused by ionizing radiation. Differences among subpopulations were also investigated at the level of chromatin organization and methylation after NotI digestion of microgel-embedded cells. In parallel experiments, the NotI-digested nucleoids were also analyzed with the use of pulsed-field gel electrophoresis (PFGE) and the DNA migration patterns were compared with the corresponding patterns from the MMCA. Significant heterogeneity in the distribution of the oxidative DNA damage, as well as intracellular variations in the NotI digestion patterns were observed in the cell population of PBMC. The combined use of both the comet assay and PFGE provides a useful model for analysis of variation in DNA damage in individual cells as well as information on size of DNA fragments.

Effects of smoking and aging on oxidative DNA damage of human lymphocytes.

The effects of H2O2-induced oxidative DNA damage in 80 healthy individuals with relation to age (20-25 and 55-60 years old) and smoking has been investigated with the comet assay technique. Both factors have shown a significant effect upon basal DNA damage with smoking appearing to have the most impact. A differentiation of the four groups response to induced oxidative damage was also observed. A distinctly separate behavior of the younger non-smokers group, when compared with the rest of the categories, was found. This is attributed to the lower degree of initial basal damage that occurs in their lymphocytes.

Study of DNA damage induction and repair capacity of fresh and cryopreserved lymphocytes exposed to H2O2 and gamma-irradiation with the alkaline comet assay.

The alkaline SCGE assay was evaluated for use with cryopreserved lymphocytes in order to obtain results similar to the freshly isolated ones. The induction of DNA damage as well as the repair capacity of gamma-rays and H2O2 exposed cryopreserved human lymphocytes was found to be the same to that of the freshly isolated. Human lymphocytes (fresh or cryopreserved) responded differently to the effects of gamma-irradiation if compared to the H2O2 treatment. The distribution of DNA damage among gamma-irradiated lymphocytes was more homogeneous compared to H2O2, both in freshly isolated and in cryopreserved cells. 2.4 micrograms/ml phytohemagglutinin at the start of a 2-h incubation in RPMI of cryopreserved samples gave similar DNA repair and distribution patterns to the 2-h post-exposure incubation of freshly isolated lymphocytes. H2O2-induced DNA damage was not repaired completely. However, the repair of gamma-rays-induced DNA damage was more efficient. These findings confirm the different mode of action of the two agents on the induction of DNA damage, as well as, the different response of the lymphocytes' DNA repair system.