The interplay of the second law of thermodynamics with the normal state and the methodology needed for the measurement of free energy of normal and malignant cells and its practical implications has not been clearly addressed in current literature. The second law of thermodynamics is one of the most fundamental laws governing the known universe at all levels. A normal cell has an exceptional ability to minimize the speed of rise in entropy to saturation of the limits of the second law. By virtue of this law, any normal resting cell is at a maximum allowable free energy. In this regard mitosis could be viewed as an attempt to maximize the lowered cellular free energy. Here we present the result of our first series of measurements, which show a significant measurable difference between the free energy of normal and malignant cells using an Olympus 510 Argon laser to calculate a diffusion correlation as well as direct visualization of motion of malignant and normal cells cultured overnight in collagen mesh. We found a significantly higher vibratory motion of the normal cells after correction for confounding factors. We also propose a new way to increase the free energy of the malignant cell to match that of its normal counterpart. This could offer hope for cure by conversion of distorted energetics of the malignant cell.

The interplay of the inflammatory microenvironment with its hypoxic niche and the potential mechanism by which they lead to malignant transformation has long been the subject of great controversy and continues to be an area of great interest today. In our previous studies we have examined this subject by using Gia2 knock out mice as the focus of our research. These mice are well known for their tendency to develop chronic inflammation in the sub mucosa of their gut, with gradual worsening and development of colon adenocarcinoma in most as they age. It has also attracted our attention that they develop a significant increase in the number of hypoxic niches in their sub mucosa, proven by EF5 staining.In contradistinction to MSI-high colon adenocarcinomas, we have also shown that histone deacetylation rather than MLH1 promoter methylation is the main mechanism of MLH1 and MSH2 deactivation in these mice. Here we show that hypoxic niches evolve under massive selective pressure of the inflammatory microenvironment as a protective shield offering survival advantage by the up regulation of NFKb and its downstream pathways, which indeed independent of true hypoxia leads to stabilization of HIF as well, securing a dual mechanism for perpetuation and expansion of hypoxic niches. We incorporated western blot and Luciferase assay of cells exposed to hypoxia+/-inflammatory cytokines to acquire data.