Modeling Excitotoxic Ischemic Brain Injury of Cerebellar Purkinje Neurons by Intravital and In Vitro Multi-photon Laser Scanning Microscopy

Author(s):  
Amanda J. Craig ◽  
Gary D. Housley ◽  
Thomas Fath
1993 ◽  
Vol 104 (4) ◽  
pp. 1175-1185 ◽  
Author(s):  
P. Buchenau ◽  
H. Saumweber ◽  
D.J. Arndt-Jovin

The regulation of DNA topology by topoisomerase II from Drosophila melanogaster has been studied extensively by biochemical methods but little is known about its roles in vivo. We have performed experiments on the inhibition of topoisomerase II in living Drosophila blastoderm embryos. We show that the enzymatic activity can be specifically disrupted by microinjection of antitopoisomerase II antibodies as well as the epipodophyllotoxin VM26, a known inhibitor of topoisomerase II in vitro. By labeling the chromatin of live embryos with tetramethylrhodamine-coupled histones, the effects of inhibition on nuclear morphology and behaviour was followed in vivo using confocal laser scanning microscopy. Both the antibodies and the drug prevented or hindered the segregation of chromatin daughter sets at the anaphase stage of mitosis. In addition, high concentrations of inhibitor interfered with the condensation of chromatin and its proper arrangement into the metaphase plate. The observed effects yielded non-functional nuclei, which were drawn into the inner yolk mass of the embryo. Concurrently, undamaged nuclei surrounding the affected region underwent compensatory division, leading to the restoration of the nuclear population, and thereby demonstrating the regulative capacity of Drosophila blastoderm embryos.


2020 ◽  
Vol 14 (01) ◽  
pp. 008-012
Author(s):  
Vinicio Hidemitsu Goto Hirai ◽  
Ricardo Machado ◽  
Maria Carolina Lucato Budziak ◽  
Lucila Piasecki ◽  
Alexandre Kowalczuck ◽  
...  

Abstract Objective This study compared different obturation techniques, analyzing percentage of areas filled with gutta-percha, sealer, and voids (PGFA, PSFA, and PVFA, respectively) in oval-shaped root canals. Materials and Methods A total of 60 extracted human mandibular central incisors were decoronated, instrumented, and irrigated using the same protocol. After drying, the root canal was filled with AH Plus labeled with 0.1% rhodamine B dye using a Lentulo spiral. The filling procedure was performed by dividing the teeth into four groups according to the respective technique: G1, cold lateral condensation; G2, continuous wave of condensation; G3, modified cold lateral condensation using an F3 master cone; and G4, modified continuous wave of condensation using an ISO (International Organization for Standardization) sized 30 gutta-percha cone. Then, slices measuring 1.5 mm in thickness were obtained 3 and 6 mm from the apex and evaluated by confocal laser scanning microscopy to determine PGFA, PSFA, and PVFA. Statistical Analysis The data were analyzed statistically with analysis of variance and Games-Howell’s tests (p = 0.05). Results The groups showed no significant differences in the apical third (3 mm from the apex). In the middle third (6 mm from the apex), G3 and G1 showed higher PGFA and PVFA, respectively. G3 showed lower PSFA than G2 and G4. Both cold techniques (G1 and G3) promoted lower PSFA than both warm techniques (G2 and G4). Conclusions Notwithstanding the limitations of this in vitro study, PGFA, PSFA, and PVFA ranged significantly only in the middle third, as observed by the different filling techniques. Higher PGFA and PVFA values were obtained for G3 and G1, respectively. Both cold techniques promoted lower PSFA than both warm techniques.


1990 ◽  
Vol 528 (1) ◽  
pp. 133-137 ◽  
Author(s):  
Roman Pohorecki ◽  
Gerald L. Becker ◽  
Pamela J. Reilly ◽  
Dennis F. Landers

PLoS ONE ◽  
2014 ◽  
Vol 9 (11) ◽  
pp. e112339 ◽  
Author(s):  
Vanessa Donega ◽  
Cora H. Nijboer ◽  
Luca Braccioli ◽  
Ineke Slaper-Cortenbach ◽  
Annemieke Kavelaars ◽  
...  

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