Micro-CT evaluation of residual material in canals filled with Activ GP or GuttaFlow following removal with NiTi instruments.

AIM: To assess the efficacy of removing Activ GP or GuttaFlow from canals using NiTi instruments. METHODOLOGY: Root canals in 55 extracted pre-molars were prepared to apical size 40, 0.04 taper. The teeth were imaged with micro-CT, and 30 teeth selected that had consistent apical size and taper of the shaped canals. They were randomly assigned to root filling with either the glass-ionomer-based ActivGP system (n = 15) or the polyvinylsiloxane-based GuttaFlow system (n = 15). After 2 weeks, canals were retreated stepwise with size 40-50 EndoSequence 0.04 taper instruments. Micro-CT scans (8 mum) were taken after use of each instrument to detect root filling residue in the coronal, middle and apical segment, and the retreatment time recorded. Residue, expressed as percentage of canal surface area, was compared between groups with t-tests, and within groups with repeated measures anova and Bonferroni-adjusted pairwise comparisons. Retreatment time was analysed with one-way anova. RESULTS: The percentage of sealer residue-coated canal surface was consistently highest (P < 0.001) in the apical third of canals, and it did not differ significantly between the two root filling groups. Stepwise enlargement from size 40 to 50 significantly decreased the amount of sealer residue in both groups (P < 0.001). Retreatment time did not differ significantly between groups. CONCLUSIONS: Both root fillings with ActivGP and GuttaFlow were removed with nickel-titanium rotary instruments. Enlargement of canals up to two sizes beyond the pre-retreatment size was necessary to minimize the amount of sealer remaining.

Plasmalogens as endogenous antioxidants: somatic cell mutants reveal the importance of the vinyl ether.

Exposure of plasmalogen-deficient variants of the murine cell line RAW 264.7 to short-term (0-100 min) treatment with electron transport inhibitors antimycin A or cyanide (chemical hypoxia) resulted in a more rapid loss of viability than in the parent strain. Results suggested that plasmalogen-deficient cells were more sensitive to reactive oxygen species (ROS) generated during chemical hypoxia; the mutants could be rescued from chemical hypoxia by using the antioxidant Trolox, an alpha-tocopherol analogue, and they were more sensitive to ROS generation by plumbagin or by rose bengal treatment coupled with irradiation. In addition, the use of buffers containing 2H2O greatly enhanced the cytotoxic effect of chemical hypoxia, suggesting the involvement of singlet oxygen. We used the unique enzymic deficiencies displayed by the mutants to differentially restore either plasmenylethanolamine (the major plasmalogen species normally found in this cell line) or its biosynthetic precursor, plasmanylethanolamine. Restoration of plasmenylethanolamine, which contains the vinyl ether, resulted in wild-type-like resistance to chemical hypoxia and ROS generators, whereas increasing levels of its precursor, which bears the saturated ether, had no effect on cell survival. These findings identify the vinyl ether double bond as a crucial element in cellular protection under these conditions and support the hypothesis that plasmalogens, through the vinyl ether, act as antioxidants to protect cells against ROS. These phospholipids might protect cells from ROS-mediated damage during events such as chemical hypoxia.

The use of DiI-marked hepatocytes to demonstrate orthotopic, intrahepatic engraftment following hepatocellular transplantation.

A novel method is described for marking primary hepatocytes with the fluorescent dye DiI prior to hepatocellular transplantation and identifying these cells within the hepatic parenchyma of recipient animals by fluorescence microscopy and flow cytometry. Optimal conditions are described for marking cells with DiI in suspension or in monolayer cultures prior to transplantation. DiI is shown to be nontoxic to hepatocytes and not to be exchanged between adjacent cells in vitro. Histological analysis of transplanted tissues shows DiI staining of engrafted hepatocytes and phagocytotic cells (Kupffer cells). This analysis shows that hepatocytes engraft within the hepatic parenchyma and exhibit a histological appearance indistinguishable from normal by conventional hematoxylin and eosin staining. Many previous reports of hepatocellular transplantation have been limited by their inability to unequivocally identify transplanted cells within the liver. These data illustrate the importance of having specific markers for transplanted cells that engraft in an orthotopic location and assume a normal morphological appearance.

Cell proliferation and DNA repair in the liver during early stages of chemical carcinogenesis.

X-chromosomal phosphoglycerate kinase mocaicisms in organ samples of female heterozygous mice provided a means to prove, because of a selective expression of one of the two allozymes, the clonal origin of carcinogen-induced preneoplastic hepatocellular populations (see also ref. 61). The occurrence of these clonal preneoplastic foci was used in rats to determine cell cycle dependent differences of transformation sensitivity and DNA repair. The highest transformation rate was found after carcinogen exposure in late G1/early S phase of the cell cycle. Experimental disturbance of DNA precursor pools by continuous thymidine infusion during carcinogen exposure results in an increased formation of preneoplastic clones. This is a further argument in favor of an essential role of base-mispairing during initiation. Cell cycle dependent fluctuations of O6-methylguanine DNA transferase with an increasing enzyme activity in late G1 and a maximum in early S phase indicate that cells possess an increased potential for eliminating promutagenic O6-methylguanine during the most transformation sensitive parts of the cell cycle to prevent base-mispairing during DNA replication or transcription. However, this putatively protective effect is limited because the enzyme is rapidly expended in the reaction and drops again in later stages of the cell cycle.