Duchenne and Becker muscular dystrophy are the major neuromuscular disorders with X-linked recessive inheritance. Preimplantation sex determination has been generally used to avoid pregnancies with these diseases. However, in order to determine if the embryo is normal, carrier or affected regardless of the sex, there is a need for a combined analysis of specific exon on dystrophin gene as well as sex determination of embryo using the same biopsied blastomere. If the exon deletion is not determinable, further diagnosis of carrier or patient can be performed by haplotype analysis. In this study, we applied the primer extension preamplification (PEP) method, which amplifies the whole genome, in 40 cases of single amniocyte and 40 cases of chorionic villus cell. We analysed haplotypes using two (CA)n dinucleotide polymorphic markers located at the end of 5' and 3' region of the dystrophin gene. Exon 46 of dystrophin gene and DYZ3 on chromosome Y were chosen as a target sequence for coamplification PCR. Upon optimizing the conditions, the amplification rates were 91.25% (73/80) for haplotypes (92.5% in amniocyte, 90% in chorionic villus cell) and 88.75% (71/80) for coamplification (85% in amniocyte, 92.5% in chorionic villus cell). The result of the study indicates that haplotypes and coamplification using PEP can be applied to prenatal and preimplantation diagnosis in Duchenne muscular dystrophy making it possible to determine if the fetus is a carrier or an affected one.
Blastomeres ; Chorionic Villi ; Diagnosis ; Dystrophin* ; Embryonic Structures ; Exons ; Fetus ; Genome ; Haplotypes* ; Humans ; Muscular Dystrophies ; Muscular Dystrophy, Duchenne ; Polymerase Chain Reaction* ; Pregnancy ; Preimplantation Diagnosis ; Wills

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'toysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.
Blastomeres ; Chorionic Villi ; Diagnosis ; DNA ; Dystrophin* ; Embryonic Structures ; Exons ; Genome ; Oocytes ; Polar Bodies ; Spermatozoa

Prenatal diagnoses were performed in 145 fetuses resulting from 73 singleton and 36 twin pregnancies, all established by intracytoplasmic sperm injection (ICS: amniocentesis in 108 patients and Chorionic villus sampling in one. The prenatal cytogenetic results were obtained from pregnancies after ICSI using ejaculated spermatozoa, epididymal spermatozoa, testicular spermatozoa and after the replacement of frozen-thawed embryos derived from ICSI. The Karyotypes were normal in 138 cases (95.2%) of the prenatal diagnoses and there were 2 cases (1.4%) de novo and 5 cases (3.4%) inherited chromosomal aberrations. The two cases of de novo abnormalities were: 46, XY, t(6;7)(q21;p22) and 47, XY, +21 (trisomy 21).
Amniocentesis ; Chorionic Villi Sampling ; Chromosome Aberrations ; Cytogenetic Analysis* ; Cytogenetics* ; Embryonic Structures ; Female ; Fetus* ; Humans ; Karyotype ; Pregnancy ; Pregnancy, Twin ; Prenatal Diagnosis ; Sperm Injections, Intracytoplasmic* ; Spermatozoa*

No abstract available.
Embryonic Structures ; Fetus ; Humans* ; Kidney*

OBJECTIVE: This study was performed to investigate the influence of coculture with human amnion cell on growth, development, implantation in mouse embryos. METHODS: Two-cell and Eight-cell stage mouse embryos were cocultured for 96 hrs with amnion cell. Embryos not treated with amnion were served control. The percentages of embryos which developed to the expanded, hatched blastocyst stage and in vitro implantation at 24hrs, 48hrs, 72hrs were determined. RESULTS: The percentages of fully expanded murine blastocysts in coculture with amnion cell were not significantly different from control. The percentages of hatched blastocysts were significantly higher in coculture group at two-cell stage (71.6%) compared to control (47.0%; p<0.05). The percentages of hatched blastocysts were significantly higher in coculture group at eight-cell stage (74.0%) compared to control (56.1%; p<0.05). In two-cell stage embryos, the percentages of implanted blastocyst in vitro were significantly higher following coculture with amnion cell (31.6%) compared to control (13.7%). CONCLUSION: Amnion cell coculture may have a stimulatory role in embryonic development, implantation compared to control. The coculture condition which induces the best effect on in vitro growth and development might be the mimics of the physiologic natural condition, just like the coculture with the tubal epithelium.
Amnion* ; Animals ; Blastocyst ; Coculture Techniques* ; Embryonic Development ; Embryonic Structures* ; Epithelium ; Female ; Growth and Development ; Humans* ; Mice* ; Pregnancy

In the rat, the neural plate appears at day 9 and the neural tube closes at day 10.3. During neurulation, the neuroepithelium is exposed directly to amniotic fluid and blood circulation begins at day 10.5. Accordingly, amniotic fluid may be an important source of nutrition for normal development of the nervous system. Among many different components of amniotic fluid, glucose is known as common currency of metabolism and the developing embryo is more dependent on this. The purpose of this study is to provide basic data on the capacity of amniotic fluid as a source of glucose for neurulating rat embryos. In the first part of this paper, isolated days 10, 11 and 16 rat embryos with intact amnion were used to pursue the change of the glucose concentratons in the amniotic fluid. The day 10 embryo amniotic fluid glucose disappeared after 20 minutes, and the day 11 amniotic fluid glucose disappeared after 33 minutes. The day 16 amniotic fluid glucose showed no significant changes during 40 minutes. In the second part of this paper, the author determined the time required for glucose concentraton in the day 10 amniotic fluid to be 0 mg% at glucose free Hanks` solution. The day 10 amniotic fluid glucose disappeared afttar 10 minutes. Another embryos were exposed to glucose free Hank`s solution for 10 minutes, and switched immediately to regular Hank`s for measuring the changes of amniotic fluid glucose, that is `charging Phenomena`. During the first 15 minutes amniotic glucose was charged to nearly normal level, and after that it decreased. These changes were similar to the results from the first experiment. These results indicate that neurulating embryo has a potential for restoring its amniotic glucose concentration to the normal level rapidly. So harmful effects of hypoglycemic states may be compensated by this `charging phenomena` of amniotic fluid during neurulation.
Amnion ; Amniotic Fluid* ; Animals ; Blood Circulation ; Embryonic Structures* ; Female ; Glucose* ; Metabolism ; Nervous System ; Neural Plate ; Neural Tube ; Neurulation ; Rats*

No abstract available.
Amino Acids* ; Blastocyst* ; Embryonic Structures*

No abstract available.
Animals ; Blastocyst* ; Embryonic Structures* ; Mice*

The patent urachus results from failure of luminal closure of the urachus. Embryologically, the urachus represents the vestigial remnant of the allantois which, in the fetus, communicates with the cloaca. The authors report a case of congenital patent urachus with review of some literatures.
Allantois ; Cloaca ; Fetus ; Phenobarbital ; Urachus*

Human embryology is the study of development from a single cell to a baby in 9 months. Implantation occurs at the end of the first week of development. The second week of development is known as the week of 2's. Gastrulation, the most characteristic event occurring in the third week, establishes three germ layers composed of ectoderm, mesoderm, and endoderm. The three germ layers and neural crest cells lead to the development of their own tissues and organs during the embryonic period, which extends from the third to the eighth week. Major congenital malformations occur in the embryonic period. The fetal period, from the third month to the day of birth, is the time for maturation of tissues and organs, and growth of the body. Because of the close relationship between embryology and congenital abnormalities, knowledge of human development is essential to assess the effects on the embryo when the mother has been exposed to teratogens. This paper briefly reviews the normal embryonic development and associated congenital malformation.
Congenital Abnormalities ; Ectoderm ; Embryology ; Embryonic Development* ; Embryonic Structures* ; Endoderm ; Female ; Gastrulation ; Germ Layers ; Human Development ; Humans* ; Mesoderm ; Mothers ; Neural Crest ; Neurulation ; Parturition ; Pregnancy ; Teratogens