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- Galluzzi, L, et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes.
- 2009
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Ingår i: Cell death and differentiation. - : Springer Science and Business Media LLC. - 1476-5403 .- 1350-9047. ; 16:8, s. 1093-107
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Forskningsöversikt (refereegranskat)abstract
- Cell death is essential for a plethora of physiological processes, and its deregulation characterizes numerous human diseases. Thus, the in-depth investigation of cell death and its mechanisms constitutes a formidable challenge for fundamental and applied biomedical research, and has tremendous implications for the development of novel therapeutic strategies. It is, therefore, of utmost importance to standardize the experimental procedures that identify dying and dead cells in cell cultures and/or in tissues, from model organisms and/or humans, in healthy and/or pathological scenarios. Thus far, dozens of methods have been proposed to quantify cell death-related parameters. However, no guidelines exist regarding their use and interpretation, and nobody has thoroughly annotated the experimental settings for which each of these techniques is most appropriate. Here, we provide a nonexhaustive comparison of methods to detect cell death with apoptotic or nonapoptotic morphologies, their advantages and pitfalls. These guidelines are intended for investigators who study cell death, as well as for reviewers who need to constructively critique scientific reports that deal with cellular demise. Given the difficulties in determining the exact number of cells that have passed the point-of-no-return of the signaling cascades leading to cell death, we emphasize the importance of performing multiple, methodologically unrelated assays to quantify dying and dead cells.
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| 2. |
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| 3. |
- Boughan, PK, et al.
(författare)
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Nucleotide-binding oligomerization domain-1 and epidermal growth factor receptor - Critical regulators of beta-defensins during helicobacter pylori infection
- 2006
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 281:17, s. 11637-11648
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Tidskriftsartikel (refereegranskat)abstract
- Host-pathogen interactions that allow Helicobacter pylori to survive and persist in the stomach of susceptible individuals remain unclear. Human beta-defensins ( hBDs), epithelial-derived antimicrobial peptides are critical components of host-defense at mucosal surfaces. The role of H. pylori-mediated NF-kappa B and epidermal growth factor receptor ( EGFR) activation on beta-defensin expression was investigated. Transient transfection studies utilizing beta-defensin promoter constructs were conducted in gastric cells with contribution of individual signaling events evaluated by the addition of specific inhibitors, small interference nucleotide-binding oligomerization domain 1( NOD1) RNA or plasmids encoding Vaccinia virus proteins that interrupt interleukin-1 and Toll-like receptor signaling. The role of individual MAPK pathways was further delineated in HEK-293 cells expressing conditional MAPK mutants. We found hBD2 expression exclusively dependent on the presence of the bacterial cag pathogenicity island, with NOD1 a critical host sensor. Impairment of murine beta-defensin 4( an orthologue of hBD2) expression in NOD1-deficient mice 7-days post-infection further confirmed the role of this cytoplasmic pattern-recognition receptor in eliciting host innate immunity. In contrast to hBD2, hBD3 expression was NOD1-independent but EGFR and ERK pathway-dependent. Importantly, Toll-like receptor signaling was not implicated in H. pylori-mediated hBD2 and hBD3 gene expression. The divergent signaling events governing hBD2 and hBD3 expression suggest temporal functional variation, such that hBD2 may contribute to antimicrobial barrier function during the inflammatory phase with hBD3 playing a greater role during the repair, wound healing phase of infection.
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| 4. |
- Nunez, Eugenia, et al.
(författare)
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Morphology, crystallization and melting of single crystals and thin films of star–branched polyesters with poly(epsilon-caprolactone) arms as revealed by atomic force microscopy.
- 2008
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Ingår i: Journal of Macromolecular Science, Part B. - : Informa UK Limited. - 0022-2348 .- 1525-609X. ; 47:3, s. 589-607
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Tidskriftsartikel (refereegranskat)abstract
- The morphology and thermal stability of different sectors in solution- and melt-grown crystals of star-branched polyesters with poly(epsilon-caprolactone) (PCL) arms, and of a reference linear PCL, have been studied by tapping-mode atomic-force microscopy (AFM). Real-time monitoring of melt-crystallization in thin films of star-branched and linear PCL has been performed using hot-stage AFM. A striated fold surface was observed in both solution- and melt-grown crystals of both star-branched and linear PCL. The presence of striations in the melt-grown crystals proved that this structure was genuine and not due to the collapse of tent-shaped crystals. The crystals of the star-branched polymers had smoother fold surfaces, which can be explained by the presence of dendritic cores close to the fold surfaces. The single crystals of linear PCL grown from solution showed earlier melting in the {100} sectors than in the {110} sectors, whereas no such sectorial dependence of the melting was found in the solution-grown crystals of the star-branched polymers. The proximity of the dendritic cores to the fold surface yields at least one amorphous PCL repeating unit next to the dendritic core and more nonadjacent and less sharp chain folding than in linear PCL single crystals; this evidently erased the difference in thermal stability between the {110} and {100} sectors. Melt-crystallization in thin polymer films at 53-55 degrees C showed 4 times faster crystal growth along b than along a, and more irregular crystals with niches on the lateral faces in star-branched PCL than in linear PCL. Crystal growth rate was strictly constant with time. Multilayer crystals with central screw dislocation (growing with or without reorientation of the b-axis) and twisting were observed in both classes of polymers.
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