The exact pathway of cell death after brain ischemia is under debate. In the present study, we used astrocytes from rat primary culture treated with persistent oxygen-glucose-deprivation (OGD) as a model of ischemia to examine the pathway of cell death and the relevant mechanisms. We observed changes in the cellular Avapritinib chemical structure morphology, the energy metabolism of astrocytes, and the percentage of apoptosis or oncosis of the astrocytes induced by OGD. Electron microscopy revealed the co-existence of ultrastructural features in both apoptosis and oncosis in individual cells. The cellular ATP
content was gradually decreased and the percentages of apoptotic and oncotic cells were increased during OGD. After 4h of OGD. ATP depletion to less than 35% of the control was observed, and oncosis became the primary pathway for astrocytic death. Increased plasma membrane permeability due to oncosis was associated with increased calpain-mediated degradation NVP-BSK805 cell line of several cytoskeletal proteins, including paxillin, vinculin, vimentin and GFAP. Pre-treatment with the calpain inhibitor 3-(4-iodophenyl)-2-mercapto-(Z)-2-propenoic acid (PD150606) could delay the OGD-induced astrocytic oncosis. These results suggest that there is a
narrow range of ATP that determines astrocytic oncotic death induced by persistent OGD and that calpain-mediated hydrolysis of the cytoskeletal-associated proteins may contribute to astrocytes oncosis. (C) 2010
Elsevier Ireland Ltd. All rights reserved.”
“Aims:
To develop a specific, fast and simple molecular method useful to detect the entomopathogenic bacterium Pseudomonas entomophila.
Methods and Results:
The use of bioinformatics tools allowed the identification of unique genes present in P. entomophila genome. Using such genes, we designed primers aiming to detect specifically P. entomophila by PCR. Furthermore, a pair of primers specifically designed to amplify the 16S rRNA gene in Pseudomonas species was used. Primer specificity was selleck chemicals checked using environmental pseudomonad and nonpseudomonad species. A 618 -bp fragment was amplified only in Pseudomonas using the 16S rDNA primers. Primers (PSEEN1497) designed to detect P. entomophila amplified a 570 -bp fragment only in P. entomophila. A duplex PCR was developed combining 16S rDNA and PSEEN1497 primers that allowed the detection of P. entomophila present in experimentally infected Drosophila melanogaster.
Conclusions:
We developed a molecular method useful to detect P. entomophila present in bacterial cultures or directly from infected insects.
Significance of the Study:
To the best of our knowledge, this is the first molecular method aiming to detect P. entomophila in environmental samples. The use of our method will facilitate studies related to ecology and insect host range of this entomopathogenic bacterium.