OBJECTIVE: The purpose of this investigation was to establish the prenatal diagnosis for identifying the risk for epidermolytic palmoplantar keratoderma(EPPK) of a fetus by sequence analysis of fetal genomic DNA from chorionic villi. METHODS: Chorionic villus sampling under transvaginal sonography at 12 weeks of gestation from a woman at risk for a child in a EPPK-affected family was perfomed. Polymerase chain reaction amplification of specific allele (PASA) assay was carried out for the detection of mutation(R162W in keratin 9 [K9] gene) previously identified in this family. Direct DNA sequencing analysis of K9 gene was accomplished to confirm the mutation. RESULTS: We had found the point mutation, R162W of K9 gene, in affected family members and confirmed by PASA assay. Affected family members were shown to have PCR products reactive with both the mutant and wildtype specific primers. Because we could not find any expected products after PASA assay with the primers la(+)/KSmt(-) of the fetal DNA, we predicted that the fetus did not inherited the mutant allele and that the fetus could be unaffected. After PASA assay, we analyzed DNA sequences of two family members to confirm the mutation. A C-to-T substitution at bp 545 was detected in the father, instead the fetus did not have any mutant band at that base pair. CONCLUSION: The PASA assay and direct DNA sequencing analysis of K9 gene through chorionic villi sampling and extraction of genomic DNA had validity to early prenatal diagnosis whether fetus was affected in EPPK or not.
Alleles ; Base Pairing ; Base Sequence ; Child ; Chorionic Villi ; Chorionic Villi Sampling ; DNA* ; Fathers ; Female ; Fetus ; Humans ; Keratin-9 ; Keratoderma, Palmoplantar, Epidermolytic* ; Point Mutation ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis* ; Sequence Analysis ; Sequence Analysis, DNA

No abstract available.
Erythrocytes* ; Polymerase Chain Reaction*

To investigate the properties and mechanism of PAF and TNF on the synthesis of prostaglandin E2 in human amnion, primary monolayer culture method was used for human amnion cell incubation. Amnion cells were incubated with various concentrations of PAF or TNF in Ca++ containing medium for various duration. Then PG E2 concentrations were measured by RIA and analyzed for the effect of PAF and TNF on PG E2 production according to their doses and incubation time. To test the role of Ca++ in E2 production, Ca++ free medium, Ca++ -channel antagonist and cyclo-oxygenase inhibitor were substituted or added in incubation medium. Following results were obtained. The synthesis of PG E2 was significantly enhanced by PAF of 10(-6) mol/L. The TNF also stimulated PG E2 synthesis at concentration of 10(-6)g/ml. The maximal level in PAF(10-6mol/L)-stimulated release of PG E2 was observed after 16 hours in incubation. The TNF(10(-6)g/ml)-induced PG E2 release was maximal after 24 hours of incubation. Combined application of PAF and TNF produced positive effect in PG E2 production. PAF or TNF stimulated-PG E2 production in Ca++ -free media was much lower than that of Ca++ -containing media. The PAF-stimulated PG E2 release was significantly inhibited by Ca++ -channel antagonist but TNF-stimulated PG E2 release was not effected by Ca++ -channel antagonist or cyclo-oxygenase inhibitor. It is strongly suggested us that both PAF and TNF enhance PG E2 release by amnion cell, although Ca++ -channel opening is essential only for PAF stimulation.
Amnion* ; Blood Platelets* ; Dinoprostone* ; Humans* ; Platelet Activating Factor* ; Prostaglandin-Endoperoxide Synthases ; Tumor Necrosis Factor-alpha*

No abstract available.
Amnion* ; Decidua* ; Female ; Humans*

No abstract available.
Ultrasonics*

No abstract available.
Female ; Fetal Blood* ; Gases* ; Labor Pain* ; Pregnancy ; Reference Values* ; Umbilical Cord*

No abstract available.
Lung*

No abstract available.
Fetal Blood* ; Umbilical Cord*

No abstract available.
Diagnosis*

No abstract available.
Female ; Gonadotropin-Releasing Hormone* ; Humans* ; Immunohistochemistry* ; In Situ Hybridization* ; Ovary* ; RNA, Messenger*