Fluorescence analysis of apoptosis induced by Pseudomonas aeruginosa in endothelial cells

Vol. 55 No. 2 Suppl., 2014
This supplement was not sponsored by Outside Organizations.


Carmen Curutiu, Mariana Carmen Chifiriuc, Florin Iordache, Coralia Bleotu, Veronica Lazar, Carmen Aurelia Mogoanta, Cristian Radu Popescu, Raluca Grigore, Serban Vifor Gabriel Bertesteanu

Intracellular invasion of professional phagocytic cells like monocytes and macrophages by a pathogen usually triggers the apoptosis of the host cells. The aim of this study was to evaluate if Pseudomonas aeruginosa, although not considered a classic intracellular pathogen, could adhere to endothelial cell surface, invade the intracellular compartment and subsequently induce apoptosis of the cells. The adherence and invasion capacity of P. aeruginosa to endothelial cells was monitored using Cravioto adapted method. The apoptotic cells were evidenced by staining with Acridine orange/Ethidium bromide. The qualitative assay of bacterial adherence to the cellular substrate revealed that all tested strains adhered to endothelial cells surface, exhibiting a diffuse, aggregative or mixed (diffuse-aggregative or localized-aggregative) pattern and 20-70% adherence rates. The adherence of P. aeruginosa induced the reorganization of cytoskeleton filaments and formation of endocytic membrane expansions. Cell free P. aeruginosa culture supernatants did not induce any cell death response, as noticed in case of whole bacterial culture, showing the capacity to induce apoptosis of endothelial cells. The fluorescence microscopy examination revealed chromatin condensation, fragmented nuclei, and membrane blebbing and apoptotic bodies in pathogen invaded cells.

Corresponding author: Raluca Grigore, MD; e-mail: raluca.grigore@3f.ro

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Roxana Folescu, Andrei Gheorghe Marius Motoc, Carmen Lacramioara Zamfir, Adrian Cosmin Ilie

Placenta, as a highly specialized organ in connecting the maternal and fetal organism, is frequently affected by a specific vascular pathology. The correlation between the macroscopic and the microscopic aspects of the fetal placenta vascularization together with the appearance of fibrin deposits is used to determine the pathological cause of death. Different studies suggest the hypothesis of precocious development of fibrin deposits during pregnancy, due to some immune-like reactions or due to the circulatory variations in the villous territory. The present research was realized on 467 placentas - out of 467 placentas collected in the past 10 years, 287 were from term births, 98 from premature births and 82 from abortions. The fibrin deposits in the placenta resulting from abortion can also be interpreted as alterations caused by the same circulatory variable conditions. In the therapeutic abortion, the placental fragments do not contain fibrin deposits. The correct approach of the macroscopic exam in the vascular pathology, accompanied by the recognition of the microscopic changes in the placenta, contributes to determine the cause of abortion, the premature birth or the term birth, followed by the death of the fetus and/or the mother. The study assesses the macroscopic and microscopic aspects of the vascular pathology at the placental level in cases with medical and legal implications. Our study also reported these placental aspects in aborted fetuses or in cases of intrauterine deaths, for various reasons considered forensic cases.

Corresponding author: Roxana Folescu, University Assistant, MD, PhD; e-mail: roxanafolescu@yahoo.com

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