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Biomedical subjects
Publications and source records attributed to D P DePaola.
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Folate deficiency during pregnancy induced by dietary deprivation or folate antagonists is teratogenic. Methotrexate is a commonly used antifolate drug that produces congenital defects, with the craniofacial complex being especially vulnerable. The molecular action of this compound, therefore, was examined in vitro using fetal rabbit palates in an organ culture system. Dihydrofolate reductase activity was measured in these palates on the 4 days preceding fusion. There was no significant fluctuation in activity with gestational age. Methotrexate treatment of the palates in vitro resulted in significant reduction of enzyme activity and also decreased total folate concentration. However, the in vitro fusion ability of these palates was not altered by exposure to methotrexate. These results suggest that since methotrexate is a potent inhibitor of DNA synthesis and has its maximal effect on rapidly dividing systems, the peak in cellular proliferative activity in the fetal rabbit palate occurs very early in its development.
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The effects of 30- and 45-minute trypsin- and pancreatin-separation on embryonic rabbit palate epithelium and mesenchyme were studied with the electron microscope. Changes in epithelium included fragmentation of the basal lamina and formation of cytoplasmic blebs associated with the basal cell layer. Changes in mesenchymal cells were first evident at 45 min of incubation and included cell fragmentation, nuclear pyknosis, and dilation of extracellular space. Results indicate a differential susceptibility of mesenchyme to the separating agent.
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Folate compounds are essential for nucleic acid metabolism and data from in vitro experiments have shown that folatee deficiency during pregnancy induced by dietary deprivation or folate antagonists is teratogenic. The craniofacial complex is a main target of this teratogen, with the lip and palate being especially vulnerable. The direct effect of folate deficiency on a target teratogenic tissue was, therefore, examined in vitro using fetal rabbit palates in an organ culture system. Folate pool sizes in the palates on the 4 days preceding fusion were determined by competitive binding assay. There was a sharp decrease in folate levels from 2.63 +/- 0.86 ng/mg wet wt on Day 15 of gestation to 1.23 +/- 0.35 ng/mg on Day 16 and 0.59 +/- 0.12 ng/mg on Day 17. The folat pool size on Day 14 was found to be quite variable with a mean of 1.93 ng/mg wet wt. Comparable results were obtained when the folate pool sizes were expressed as nanograms of folate per milligram of protein. The effect of a folate-deficiency culture medium on in vitro fusion and folate pool sizes was examined. The results of incubating palatal shelves excised on Days 14, 15, 16, and 17 of gestation for 5, 4, 3, and 2 days, respectively, show that a folate-depleted medium neither prevented nor delayed fusion. An examination of the palatal pool size after incubation in folate-deficient media revealed that while there was some decrease in folate levels, the palates apparently retained sufficient folate to support cellular activity.
Palatal shelves from 14-day-old embryonic mice were exposed to a 27.12 MHz pulsed non-ionizing radiofrequency (Diapulse) for 20 min followed by 24-hour organ culture in nutrient agar. Diapulse-treated palatal shelves showed induction of cartilage within the mesenchymal compartment and loss of the overlying epithelium in contrast to controls which were free of cartilage. The results are thought to be independent of thermal changes and may be due to calcium flux within the tissue.
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