( ...)Quantification of Nucleosomes in Serum,,,by the Cell Death Detection ELISAplus

Cell death as a counterpart to cell proliferation plays an essential role in the homeostasis of cell numbers in adult organisms, furthermore in eliminating cells damaged by irradiation, chemotherapeutic drugs, hyperthermia and other influences [1-3]. There are two distinct ways of eukaryotic cell death: oncosis and apoptosis. How cells die does not so much depend on the type of the stimulus, as on dose and severity of a lesion, as well as on cell type [4-6]. During apoptosis, various endonucleases are activated which cleave the chromatin into nucleosomesized fragments of 180 bp and multiples thereof, generating the typical ladder pattern observed with agarose gel electrophoresis [1-8].
Nucleosomes are elementary units of chromatin formed by an octamer of histones and 146 bp DNA wrapped around it. The nucleosomes are connected by linker DNA of about 15 -100bp which are the preferential binding sites of the endonucleases [9-11]. Under physiological conditions, nucleosomes are packed into apoptotic bodies and phagocytized by macrophages or neighboring cells [1-4]. In situations of enhanced cell death these mechanisms are overloaded and nucleosomes are also released into circulation [12-15]. Quantifying these circulating nucleosomes may offer the fascinating possibility of estimating the extent of cell death wherever it happens. This could be especially useful for follow up studies and for monitoring individuals during therapy as the material is obtained easily by venipuncture.

The Cell Death Detection ELISAplus (CDDE) was developed as a cellular test to detect nucleosomes in cytoplasm prior to disintegration of the plasma membrane which is a well-known hallmark of apoptosis [1-3, 7, 8]. We adapted the CDDE for its use in serum samples and im
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