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Some difficult microorganisms, including Chlamydophila pneumoniae, are associated with the atherosclerotic tissue damage.The aim of this study was to evaluate the employment of culture together molecular techniques in order to define the possible role of C. pneumoniae in the atherosclerotic tissue damage. Atheromatous carotid plaques (ACP) were obtained by endoarterectomies from 10 patients with severe stenosis of the internal carotid artery. Each specimen was divided in three parts: a proximal tract to heart, without stenosis, a medial tract, corresponding to the atheromatous plaque, and a distal tract, above the plaque.Aliquots were employed to perform cultures for C. pneumoniae on Hep - 2 cell line in DMEM. DNA and total RNA were extracted from 50-70 mg. of tissue sample and from Hep - 2 106 cultures to investigate 16S rRNA, momp and hsp60 genes.The PCR and RT-PCR resulted negative for momp gene of C. pneumoniae in all samples. PCR and RT-PCR resulted positive for 16S rRNA or hsp60 genes of C. pneumoniae in the proximal portion of two ACPs with hemorrhagic evolution in two patients, one of which complicated with a retinal tromboembolic outcome. Molecular analyses on C. pneumoniae growing from the cultures are in progress.The DNA and RNA amplification of different portions from ACP seems to be useful to evidence the effective localization of C. pneumoniae in the atheromatous arterial tissue. The highly gene expression of C. pneumoniae hsp60 in a patient with acute hemorrhagic evolution of the carotid plaque may suggest that C. pneumoniae might partecipate in the atherogenesis and induce atherosclerosis complications by inflammatory pathways (activation of cytokines, endothelial factors and matrix-degrading metalloproteinases).