The function of NF _B in the regulation of apoptosis is questionable but it has demonstrated an ability that in several cell lines NF _B mediates apoptosis and that its activity is down regulated by the over expression of GW0742. These studies suggest that apoptosis could intervene in the results obtained under hypo osmotic conditions that might be induced via the induction of NF _B. Further studies must evaluate correctly the result of CTLL 2 cells in a hypo osmolar environment. It is significant that you will find very few studies that build hypo osmolality, but it confirms the necessity to use a hypotonic solution to reduce the test substance particularly if used at low concentration. The results of low pH conditions are very different in line with the cell type. In CTLL 2 and CTLL 2 Bcl2 cells, minimal pH induces genotoxicity without major apoptosis. It was claimed that low pH can induce DNA damage in CHO K1 cells both in the absence and in the clear presence of S9 mix. On the other hand, no DNA damage was observed within a 4 h treatment at pH 2. 73 in rat lymphocytes or during a 1 h treatment at pH 6. 0 in human lymphocytes. However, these different email address details are presumably due not just to differences in the cell system used but also to the period of the therapy at the lower pH. This indicates the requirement for standardizing the method used for the in vitro micronucleus examination in CTLL 2 and CTLL 2 Bcl2 cells. More over, it appears that a small Organism loss of intracellular pH inhibits protein synthesis and progression through the G1 phase of Chinese hamster fibroblast cells. Thus, destabilization of DNA, including string breakage, could take place consequently of dehistonization of chromatin due to the inhibition of protein synthesis. Under alkaline pH conditions, we observed the look of apoptosis concomitant by having an increase of the amount of micronucleated CTLL 2 cells. On one other hand, Morita et al. Discovered that strong simple pH in the presence of rat liver S9 increases the clastogenic action of S9 or triggers new clastogens by break down of the S9. Neither apoptosis or chromosome aberration was seen in transfected cells. Apoptosis is apparently the main factor affecting the outcome CAL-101 PI3K inhibitor obtained under alkaline conditions. Alkaline conditions at pH 8. 5 simultaneously induced apoptosis and look of micronucleated cells. In the last step, to assess the maximal pH range to perform the assay, we employed a range of pH around the normal values. These results suggested that optimal culture conditions are between pHs 6. 7 and 7. 7. In summary, we proved that the clastogenic a reaction to extreme culture conditions is likely to yield unreliable outcomes, and we demonstrated that apoptosis and necrosis may restrict the induction of micronuclei in in vitro tests.