At present, animal experimentation remains central to our understanding of human disease-related processes and of the biological effects of many substances. Traditional experiments have relied heavily on invasive techniques to monitor changes in blood biochemistry, tissue structure or function, or to phenotype or genotype genetically modified animals. In some cases, a proportion or all of the animals used during the course of a study may be sacrificed for histopathological assessment. In most cases, this is to track the progression or regression of a disease over time, or to determine the levels of toxicity evident in specific organs or tissues. However, many of these techniques fail to provide details of how a disease develops or how a substance elicits its effects. In recent years there has been a gradual increase in the application of imaging techniques that were originally developed and used in fundamental research or in medicine. These non-invasive techniques allow diseases, and responses to exogenous substances, to be monitored in a temporal and spatial manner, therefore allowing a greater amount of information to be derived from smaller numbers of animals, which in turn, increases the statistical validity of the data by reducing the level of experimental variation. Non-invasive imaging also allows more informative and humane endpoints to be used and, perhaps most importantly, allows functional details to be studied in the context of a living animal. Some of the recent developments within the field of non-invasive imaging and their significance with respect to animal welfare and the understanding of human physiology are discussed.

Epigenetic changes are inherited alterations in DNA that affect gene expression and function without altering the DNA sequence. DNA methylation is one epigenetic process implicated in human disease that is influenced by diet. DNA methylation involves addition of a 1-C moiety to cytosine groups in DNA. Methylated genes are not transcribed or are transcribed at a reduced rate. Global under-methylation (hypomethylation) and site-specific over-methylation (hypermethylation) are common features of human tumours. DNA hypomethylation, leading to increased expression of specific proto-oncogenes (e.g. genes involved in proliferation or metastasis) can increase the risk of cancer as can hypermethylation and reduced expression of tumour suppressor (TS) genes (e.g. DNA repair genes). DNA methyltransferases (DNMT), together with the methyl donor S-adenosylmethionine (SAM), facilitate DNA methylation. Abnormal DNA methylation is implicated not only in the development of human cancer but also in CVD. Polyphenols, a group of phytochemicals consumed in significant amounts in the human diet, effect risk of cancer. Flavonoids from tea, soft fruits and soya are potent inhibitors of DNMT in vitro, capable of reversing hypermethylation and reactivating TS genes. Folates, a group of water-soluble B vitamins found in high concentration in green leafy vegetables, regulate DNA methylation through their ability to generate SAM. People who habitually consume the lowest level of folate or with the lowest blood folate concentrations have a significantly increased risk of developing several cancers and CVD. This review describes how flavonoids and folates in the human diet alter DNA methylation and may modify the risk of human colon cancer and CVD.