Interleukin 6 (IL6) plays key roles in hematopoiesis, immune, and acute phase responses. Dysregulated IL6 expression is implicated in diseases such as atherosclerosis and arthritis. We have examined the functional effect of four polymorphisms in the IL6 promoter (؊597G3 A, ؊572G3 C, ؊373A n T n , ؊174G3 C) by identifying the naturally occurring haplotypes and comparing their effects on reporter gene expression. The results indicate different transcriptional regulation in the ECV304 cell line compared with the HeLa cell line, suggesting cell type-specific regulation of IL6 expression. The haplotypes showed functional differences in the ECV304 cell line; transcription was higher from the GG9/ 11G haplotype and lower from the AG8/12G allele. The differences suggest that more than one of the polymorphic sites is functional; the base differences at distinct polymorphic sites do not act independently of one another, and one polymorphism influences the functional effect of variation at other polymorphic sites. These results show that genetic polymorphisms in the promoter influence IL6 transcription not by a simple additive mechanism but rather through complex interactions determined by the haplotype. IL6,1 a multifunctional cytokine with a central role in host defense (1-3), has diverse functions including stimulation of the hepatic acute phase response to infection and injury (4), differentiation and/or activation of macrophages and T cells, growth and terminal differentiation of B cells, support of multipotential colony formation by hematopoietic stem cells, and neural differentiation. IL6 is not constitutively expressed but is highly inducible and is produced in response to a number of inflammatory stimuli such as IL1, platelet-derived growth factor, tumor necrosis factor ␣ (TNF␣), bacterial products such as endotoxin, and viral infection. Glucocortocoids produced as part of the inflammatory response act to enhance some IL6 effects, such as acute phase protein synthesis, but down-regulate IL6 expression, providing a negative feedback pathway on the inflammatory response in vivo. Many cell types produce IL6 in response to noxious stimuli, including monocytes/macrophages (5), fibroblasts (6), endothelial cells (7), adipocytes (8), T cells (9), and mast cells (10).IL6 is an essential mediator of the acute phase response. In IL6 knockout mice, the T cell-dependent antibody response is dramatically compromised in response to localized infection and tissue damage, with impairment of the response to certain viral infections; macrophage stimulation was also deficient (11-13). IL6-deficient mice also show 20 -40% reduced numbers of thymocytes and peripheral T cells, suggesting the involvement of IL6 in T cell proliferation. Dysregulated IL6 production is implicated in the pathology of several disease processes. Constitutively high levels of IL6 in transgenic murine B cell lineages results in fatal plasmacytosis (14) and has been implicated in human multiple myeloma (15-17) and Kaposi's sarcoma (18). Increased IL6 levels are ...

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A comparative cross platform evaluation of real-time polymerase chain reaction detection of DNA sequences present in Roundup Ready soya was undertaken using the ABI 7700 and Roche Lightcycler detection systems in combination with 3 different detection chemistries: TaqMan, Scorpion primers, and SYBR Green I fluorescent dye. Various copy numbers of a plasmid containing the soya lectin sequence were used to determine the sensitivity and reproducibility of the different technology combinations and to examine both inter and intra machine variability. To examine the relative accuracy of each technology, the genetically modified soya content of baked products containing known amounts of Roundup Ready soya was determined by detection of lectin and the EPSPS transgene. It was determined that the combination of TaqMan detection chemistry and the ABI 7700 platform represented the best method for quantitative detection of genetically modified organisms in terms of both precision and accuracy.

The detection of genetically modified crops in foodstuff relies on detection of transgenic DNA or protein material in the sample matrix. Purified DNA or proteins are used as analytical material for polymerase chain reaction technologies and immunodiagnostics. Successful sample preparation is critical to the validity of subsequent analysis. For routine analysis, a good sample preparation technique should be simple, safe, and inexpensive while reproducibly generating DNA/protein of sufficient quality and yield. The suitability of isolated DNA or protein as an analyte for a detection or characterization technique depends on amount or concentration, purity, and integrity, each of which may be influenced by sample matrix and the extraction technique, and, in turn, may impact the validity of analytical techniques. The key sample preparation steps of homogenization, pretreatment, extraction, and purification are discussed as well as typical analytical methods. Consideration is given to application of these steps for particular sample matrixes to maximize yield, reduce inhibition effects, and minimize contamination. The choice of the most appropriate and valid methods for sample preparation from particular foods is discussed with respect to DNA analysis. Attention is also given to ease of use, cost, and generic applicability of the procedures.