We have investigated the cell death of a Chinese hamster ovary mutant (MT-58) with a thermo-sensitive CTP:phosphocholine cytidylyltransferase, the rate-limiting enzyme of the CDP-choline pathway for phosphatidylcholine biosynthesis (Esko, J. D., Wermuth, M. M., and Raetz, C. R. H. (1981) J. Biol. Chem. 256, 7388-7393). After MT-58 cells were shifted to the restrictive temperature of 40 degrees C, the cytidylyltransferase was inactivated immediately leading to a decrease in phosphatidylcholine biosynthesis and cell death. DNA content and number of cells in the S phase decreased significantly in the dying MT-58 cells according to flow cytometrical analyses. The fragmentation of genomic DNA was detected by DNA ladders in agarose gel and release of the prelabeled genomic DNA into cytosolic fractions 14 h after the temperature shift. The dying cells underwent a dramatic reduction of cellular volume while maintaining the membrane containment of cellular contents. These events indicated that the inactivation of cytidylyltransferase triggered apoptosis in Chinese hamster ovary cells. This is the first report that apoptosis was induced in cultured cells, not by an added agent, but by a mutation in phospholipid biosynthesis.

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These results indicate that in vivo TBA results in a degree of functional impairment of vascular smooth muscle that persists for at least 7 days. This result is consistent with previous observations of the acute effects of TBA in isolated arteries. Furthermore, these results support the hypothesis that normal smooth muscle function is required for the development of vasospasm. Finally, these results indicate that TBA performed before the onset of vasospasm prevents its development.

ObjectTo better understand the fate of islet isografts and allografts, we utilized a magnetic resonance (MR) imaging technique to monitor mouse islets labeled with a novel MR contrast agent, chitosan-coated superparamagnetic iron oxide (CSPIO) nanoparticles.Materials and MethodsAfter being incubated with and without CSPIO (10 µg/ml), C57BL/6 mouse islets were examined under transmission electron microscope (TEM) and their insulin secretion was measured. Cytotoxicity was examined in α (αTC1) and β (NIT-1 and βTC) cell lines as well as islets. C57BL/6 mice were used as donors and inbred C57BL/6 and Balb/c mice were used as recipients of islet transplantation. Three hundred islets were transplanted under the left kidney capsule of each mouse and then MR was performed in the recipients periodically. At the end of study, the islet graft was removed for histology and TEM studies.ResultsAfter incubation of mouse islets with CSPIO (10 µg/mL), TEM showed CSPIO in endocytotic vesicles of α- and β-cells at 8 h. Incubation with CSPIO did not affect insulin secretion from islets and death rates of αTC1, NIT-1 and βTC cell lines as well as islets. After syngeneic and allogeneic transplantation, grafts of CSPIO-labeled islets were visualized on MR scans as persistent hypointense areas. At 8 weeks after syngeneic transplantation and 31 days after allogeneic transplantation, histology of CSPIO-labeled islet grafts showed colocalized insulin and iron staining in the same areas but the size of allografts decreased with time. TEM with elementary iron mapping demonstrated CSPIO distributed in the cytoplasm of islet cells, which maintained intact ultrastructure.ConclusionOur results indicate that after syngeneic and allogeneic transplantation, islets labeled with CSPIO nanoparticles can be effectively and safely imaged by MR.

Purple rice bran contains high amounts of natural antioxidants that consist of water- and lipid-soluble compounds. Hexane-insoluble and hexane-soluble fractions were separated from 100% methanolic extract from purple rice bran (RBE-HI and RBE-HS, respectively). Total anthocyanin, tannin, flavonoid, and phenolics contents were determined in those fractions, and their antioxidant capacities were evaluated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capability, oxygen radical absorbance capacity (ORAC), and iron chelating capability (ICC). RBE-HI and RBE-HS were also added to restructured patties made from minced channel catfish (Ictalurus punctatus) belly flap meat. Lipid oxidation, color, and/or textural properties were determined for raw and cooked patties during a 12-d storage at 4 degrees C. All antioxidant indices, except for ICC, of RBE-HI were significantly higher than those of RBE-HS due probably to its higher anthocyanin content (P < 0.05). RBE-HS showed higher ICC (P < 0.05). However, both fractions showed similar antioxidant activity in raw and cooked patties during storage, resulting from the complexity of antioxidant action in food systems. Textural properties (hardness, cohesiveness, chewiness, and springiness) in cooked patties with RBE-HS and RBE-HI were well maintained during storage, but changed significantly in the control (P < 0.05). Only RBE-HS limited microbial growth in raw patties during storage (P < 0.05), but its inhibitory effect was marginal because of low-dose and physical interactions with the matrix. L* (lightness) and a* (redness) of raw and cooked patties decreased significantly by both fractions, whereas b* (yellowness) was significantly decreased by RBE-HI and increased by RBE-HS (P < 0.05). In conclusion, we suggest that purple rice bran extract is applicable to meat products as a natural preservative, but color change in the products may limit its application.