OBJECTIVE: To determine whether the serum beta-human chorionic gonadotropin (hCG) profile following preimplantation genetic diagnosis (PGD) is lower than that of intracytoplasmic sperm injection (ICSI) cycles. METHODS: A total of 129 PGD cycles and 1,161 age-matched ICSI cycles, which resulted in pregnancy (serum beta-hCG> or =5 mIU/mL) on post-ovulation day (POD) 12 were included. We compared the mean serum beta-hCG levels on POD 12, 14, 21, and 28, doubling time of serum hCG, and created a cut-off value for predicting a singleton pregnancy in each group. RESULTS: The mean serum beta-hCG concentration of the PGD group was significantly lower than that of the control group on POD 12, 14, and 21. The doubling time of serum beta-hCG at each time interval showed no significant difference. The cut-off-value of serum beta-hCG for predicting a single viable pregnancy was 32.5 mIU/mL on POD 12 and 113.5 mIU/mL on POD 14 for the PGD group, which was lower than that for the control group. CONCLUSION: Blastomere biopsy may decrease the beta-hCG-producing activity of the trophoblasts, especially in early pregnancy. Setting a lower cut-off value of serum beta-hCG for predicting pregnancy outcomes in PGD may be needed.
Biopsy ; Blastomeres ; Chorionic Gonadotropin ; Female ; Humans ; Pregnancy ; Pregnancy Outcome ; Preimplantation Diagnosis ; Prostaglandins D ; Sperm Injections, Intracytoplasmic ; Trophoblasts

OBJECTIVE: To determine whether the serum beta-human chorionic gonadotropin (hCG) profile following preimplantation genetic diagnosis (PGD) is lower than that of intracytoplasmic sperm injection (ICSI) cycles. METHODS: A total of 129 PGD cycles and 1,161 age-matched ICSI cycles, which resulted in pregnancy (serum beta-hCG> or =5 mIU/mL) on post-ovulation day (POD) 12 were included. We compared the mean serum beta-hCG levels on POD 12, 14, 21, and 28, doubling time of serum hCG, and created a cut-off value for predicting a singleton pregnancy in each group. RESULTS: The mean serum beta-hCG concentration of the PGD group was significantly lower than that of the control group on POD 12, 14, and 21. The doubling time of serum beta-hCG at each time interval showed no significant difference. The cut-off-value of serum beta-hCG for predicting a single viable pregnancy was 32.5 mIU/mL on POD 12 and 113.5 mIU/mL on POD 14 for the PGD group, which was lower than that for the control group. CONCLUSION: Blastomere biopsy may decrease the beta-hCG-producing activity of the trophoblasts, especially in early pregnancy. Setting a lower cut-off value of serum beta-hCG for predicting pregnancy outcomes in PGD may be needed.
Biopsy ; Blastomeres ; Chorionic Gonadotropin ; Female ; Humans ; Pregnancy ; Pregnancy Outcome ; Preimplantation Diagnosis ; Prostaglandins D ; Sperm Injections, Intracytoplasmic ; Trophoblasts

The aim of this study was to investigate whether stereotactic body radiotherapy (SBRT) can salvage gastric cancer patients with para-aortic lymph node (PALN) recurrence. From January 2003 to December 2006, 7 patients were treated for isolated PALN recurrence from gastric cancer after curative resection. Follow up durations ranged from 19 to 33 months (median; 26 months), and SBRT doses from 45 Gy to 51 Gy (median 48 Gy) in 3 fractions. Disease progression-free and overall survivals and toxicities were recorded. Response to treatment was assessed by computed tomography. Final patient outcomes were as follows: 2 were alive without evidence of disease, 3 remained alive with disease, and 2 patients died of disease. Five of 7 patients showed complete response and 2 patients partial response between 3 and 11 months after SBRT. Three-year overall and disease progression-free survival rates post-SBRT were 43% and 29%, respectively. No severe complication was detected during follow-up. Selected patients with isolated PALN recurrence can be salvaged by SBRT without severe complications.
Adult ; Aged ; Disease-Free Survival ; Female ; Humans ; Lymph Nodes/surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local/*radiotherapy/surgery ; Radiosurgery ; Radiotherapy Dosage ; Recurrence ; Stomach Neoplasms/diagnosis/pathology/*surgery ; Survival Analysis

The aim of this study was to investigate whether stereotactic body radiotherapy (SBRT) can salvage gastric cancer patients with para-aortic lymph node (PALN) recurrence. From January 2003 to December 2006, 7 patients were treated for isolated PALN recurrence from gastric cancer after curative resection. Follow up durations ranged from 19 to 33 months (median; 26 months), and SBRT doses from 45 Gy to 51 Gy (median 48 Gy) in 3 fractions. Disease progression-free and overall survivals and toxicities were recorded. Response to treatment was assessed by computed tomography. Final patient outcomes were as follows: 2 were alive without evidence of disease, 3 remained alive with disease, and 2 patients died of disease. Five of 7 patients showed complete response and 2 patients partial response between 3 and 11 months after SBRT. Three-year overall and disease progression-free survival rates post-SBRT were 43% and 29%, respectively. No severe complication was detected during follow-up. Selected patients with isolated PALN recurrence can be salvaged by SBRT without severe complications.
Adult ; Aged ; Disease-Free Survival ; Female ; Humans ; Lymph Nodes/surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local/*radiotherapy/surgery ; Radiosurgery ; Radiotherapy Dosage ; Recurrence ; Stomach Neoplasms/diagnosis/pathology/*surgery ; Survival Analysis

Ornithine transcarbamylase (OTC) deficiency is an X-linked co-dominant disorder. A couple, with a previous history of a neonatal death and a therapeutical termination due to OTC deficiency, was referred to our center for preimplantation genetic diagnosis (PGD). The female partner has a nonsense mutation in the exon 9 of the OTC gene (R320X). We carried out nested polymerase chain reaction (PCR) for R320X mutation and fluorescence in situ hybridization (FISH) for aneuploidy screening. Among a total of 11 embryos, two blastomeres per embryo from 9 embryos were biopsied and analyzed by duplex-nested PCR and FISH, and one blastomere per embryo from 2 embryos by only duplex-nested PCR. As a result of PCR and restriction fragment length polymorphism analysis, four embryos were diagnosed as unaffected embryos having the normal OTC gene. Among these embryos, only one embryo was confirmed as euploidy for chromosome X, Y and 18 by FISH analysis. A single normal embryo was transferred to the mother, yielding an unaffected pregnancy and birth of a healthy boy. Based on our results, PCR for mutation loci and FISH for aneuploidy screening with two blastomeres from an embryo could provide higher accuracy for the selection of genetically and chromosomally normal embryos in the PGD for single gene defects.
Adult ; Aneuploidy ; Codon, Nonsense ; DNA Primers ; Exons ; Female ; Humans ; In Situ Hybridization, Fluorescence/*methods ; Infant, Newborn ; Male ; Ornithine Carbamoyltransferase/*deficiency ; Polymerase Chain Reaction/*methods ; Pregnancy ; Pregnancy Outcome ; Preimplantation Diagnosis/*methods

The pre-diagnostic test for preimplantation genetic diagnosis (PGD) of long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency was performed by polymerase chain reaction (PCR) and direct sequencing for hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A hydratase (HADHA) gene. We obtained unexpected genotyping results of HADHA gene by allele drop-out in the analysis of patients' genomic DNA samples with a referred PCR primer set. Upon further analysis with a re-designed primer set, we found a novel single nucleotide polymorphism (SNP) at the referred primer-binding site in the normal allele of HADHA gene (NT_022184, 5233296 a>t). We found that the frequency of this novel SNP was 0.064 in Korean population. Pre-diagnostic test using single lymphocytes and clinical PGD were successfully performed with the re-designed primer set. Nineteen embryos (95.0%) among 20 were successfully diagnosed to 5 homozygous mutated, 8 heterozygous carrier and 6 wild type. Among 6 normal embryos, well developed and selected 4 embryos were transferred into the mother's uterus, but a pregnancy was not achieved. We proposed that an unknown SNP at primer-binding sites would be a major cause of allele drop-out in the PGD for single gene dis-order.
*Preimplantation Diagnosis ; *Polymorphism, Single Nucleotide ; Polymerase Chain Reaction ; Mutation ; Multienzyme Complexes/*genetics ; Male ; Humans ; Female ; Binding Sites ; Adult ; 3-Hydroxyacyl CoA Dehydrogenases/deficiency

OBJECTIVES: The risk of aneuploidies of embryos increases in advanced maternal age or parental karyotype abnormality and it results in poor reproductive outcomes such as recurrent spontaneous abortion (RSA) or repeated implantation failure (RIF). Preimplantation genetic diagnosis for aneuploidy screening (PGD-AS) can be applied for better ART outcome by selecting chromosomally normal embryos. The aim of this study is to evaluate the clinical outcome of PGD-AS and which group can get much benefit from PGD-AS among the patients expected to have poor reproductive outcome. METHODS: In 42 patients, 77 PGD cycles were performed for aneuploidy screening. Patients were allocated to 3 groups according to the indication of PGD-AS: group I-patients with old age (> or =37) and RIF more than 3 times (n=11, mean age=42.2 yrs.), group II-patients with RSA (> or = 3 times) associated with aneuploid pregnancy (n=19, mean age=38.9 yrs.), group III-parental sex chromosome abnormality or mosaicism (n=18, mean age=29.6 yrs.) including Turner syndrome, Klinefelter syndrome and 47,XYY. PGD was performed by using FISH for chromosome 13, 16, 18, 21, X and Y in group I and II, and chromosome X, Y and 18 (or 17) in group III. RESULTS: Blastomere biopsy was successful in 530 embryos and FISH efficiency was 92.3%. The proportions of transferable embryos in each group were 32.5+/-17.5%, 23.0+/-21.7% and 52.6+/-29.2% (mean +/- SD), respectively, showing higher normal rate in group III (group II vs. III, p<0.05). The numbers of transferred embryos in each group were 3.9+/-1.5, 1.9+/-1.1 and 3.1+/-1.4 (mean +/- SD), respectively. The clinical pregnancy rates per transfer was 0%, 30.0% and 20.0%, and it was significantly higher in group II (group I vs. group II, p<0.05). The overall pregnancy rate per transfer was 19.6% (10/51) and the spontaneous abortion rate was 20% (2/10) of which karyotypes were euploid. Nine healthy babies (one twin pregnancy) were born with normal karyotype confirmed on amniocentesis. CONCLUSION: Our data suggests that PGD-AS provides advantages in patients with RSA associated with aneuploidy or sex chromosome abnormality, decreasing abortion rate and increasing ongoing pregnancy rate. It is not likely to be beneficial in RIF group due to other detrimental factors involved in implantation.
Abortion, Induced ; Abortion, Spontaneous ; Amniocentesis ; Aneuploidy* ; Biopsy ; Blastomeres ; Chromosomes, Human, Pair 13 ; Embryonic Structures ; Female ; Humans ; Karyotype ; Klinefelter Syndrome ; Mass Screening* ; Maternal Age ; Mosaicism ; Parents ; Pregnancy ; Pregnancy Rate ; Preimplantation Diagnosis* ; Prostaglandins D ; Sex Chromosome Aberrations ; Turner Syndrome ; Twins

OBJECTIVE: Preimplantation genetic diagnosis (PGD) is reserved for couples with a risk of transmitting a serious and incurable disease, and hence avoids the undesirable therapeutic abortion. In this study, we evaluated the efficacy of PGD for Duchenne muscular dystrophy (DMD) cases by the fluorescent PCR with polymorphic linked markers and the conventional duplex-nested PCR methods. METHODS: Biopsy of one or two blastomeres was done from the embryos fertilized by ICSI on the third day after fertilization. We performed two cases of PGD-DMD by the duplex-nested PCR for the causative mutation loci and the SRY gene on Y chromosome. The triplex fluorescent PCR for the mutation loci, the SRY gene and the polymorphic microsatellite marker on X chromosome was applied for two cases of PGD-DMD. RESULTS: By the duplex-nested PCR, successful diagnosis rate was 95.5% (21/22), but we could not discriminate the female embryos whether normal or carrier in this X-linked recessive disease. However, the triplex fluorescent PCR method showed 100% (27/27) of successful diagnosis rate, and all female embryos (n=17) were distinguished normal (n=10) from carrier (n=7) embryos. Unaffected and normal embryos were transferred into mother's uterus after diagnosis. A healthy normal male was achieved after PGD with the duplex-nested PCR method and a twin, a male and a female, were delivered with triplex fluorescent PCR method. The normality of dystrophin gene was confirmed by amniocentesis and postnatal genetic analysis in all offsprings. CONCLUSION: The fluorescent PCR with polymorphic marker might be useful in improving the specificity and reliability of PGD for single gene disorders.
Abortion, Therapeutic ; Amniocentesis ; Biopsy ; Blastomeres ; Diagnosis ; Dystrophin ; Embryonic Structures ; Family Characteristics ; Female ; Fertilization ; Genes, sry ; Humans ; Male ; Microsatellite Repeats ; Muscular Dystrophy, Duchenne* ; Polymerase Chain Reaction* ; Pregnancy ; Preimplantation Diagnosis* ; Prostaglandins D ; Sensitivity and Specificity ; Sperm Injections, Intracytoplasmic ; Twins ; Uterus ; X Chromosome ; Y Chromosome

No abstract available.
Epidermolysis Bullosa ; Muscular Dystrophy, Duchenne ; Ornithine Carbamoyltransferase ; Polymerase Chain Reaction* ; Preimplantation Diagnosis*

OBJECTIVE: Preimplantation genetic diagnosis (PGD) is reserved for couples with a risk of transmitting a serious and incurable disease, and hence avoids the undesirable therapeutic abortion. Herein, we report the result of PGD to carriers at risk of transmitting ornithine transcarbamylase (OTC) deficiency, junctional epidermolysis bullosa (EB) and lactic acidosis (LA) due to defect of pyruvate dehydrogenase alpha1 gene, respectively. METHODS: The ovarian stimulation, oocyte retrieval and ICSI procedure were undergone by conventional protocols. PGD for single gene disorders was carried out after biopsy of one or two blastomeres from the embryos on the third day. We performed the duplex nested PCR of the simultaneous amplification for the causative mutation loci as well as the SRY gene on Y chromosome in case of OTC deficiency and LA. Two different mutation loci of ITGB4 gene in EB case were amplified by the same protocol. The PCR products were analyzed by agarose gel electrophoresis, restriction fragment length polymorphism analysis or direct DNA sequencing. RESULTS: A total of 26 embryos were analyzed by duplex nested PCR. One or two blastomeres were biopsied, and successful diagnosis rate of PGD with PCR was 92.3% (24/26). There was no contamination in all PCR samples of negative controls (n=67). Five embryos (19.2%) were diagnosed as normal embryos, which were transferred to the mothers' uterus in each cases. In OTC deficiency case, singleton pregnancy was established. At 17 weeks of gestation, genetic normality of OTC gene in fetus was confirmed by amniocentesis. A healthy baby was successfully delivered at 36 weeks of gestation in OTC deficiency case. Unfortunately, pregnancies were not achievement in cases of EB and LA. CONCLUSION: This is the first report in Korea that healthy baby was born after specific PGD for OTC deficiency. Our results demonstrate that duplex nested PCR for single cell is an efficient method in identifying the gender and single gene mutation or two different mutation loci, simultaneously. This PGD procedure could provide normal healthy baby to the couple with a high risk of transmitting genetic diseases.
Abortion, Therapeutic ; Acidosis, Lactic* ; Amniocentesis ; Biopsy ; Blastomeres ; Diagnosis ; Electrophoresis, Agar Gel ; Embryonic Structures ; Epidermolysis Bullosa* ; Epidermolysis Bullosa, Junctional ; Family Characteristics ; Female ; Fetus ; Genes, sry ; Korea ; Oocyte Retrieval ; Ornithine Carbamoyltransferase Deficiency Disease* ; Ornithine Carbamoyltransferase* ; Ornithine* ; Ovulation Induction ; Oxidoreductases ; Polymerase Chain Reaction* ; Polymorphism, Restriction Fragment Length ; Pregnancy ; Preimplantation Diagnosis* ; Prostaglandins D ; Pyruvic Acid ; Sequence Analysis, DNA ; Sperm Injections, Intracytoplasmic ; Uterus ; Y Chromosome