To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group. Design: Prospective study. Setting: Genome Research Center for Reproductive Medicine and Infertility...To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group. Design: Prospective study. Setting: Genome Research Center for Reproductive Medicine and Infertility, Korea Ministry of Health & Welfare. Patient(s): Thrity patients with POF and 30 control individuals. Interventions(s): The mitochondrial DNA content was quantified using real-time PCR. The effectiveness of the assay was determined by relative quantification using the comparative threshold cycle (CT) method. Main Outcome Measure(s): Relative quantification of mitochondrial DNA content. Interventions(s): The mitochondrial DNA content was significantly lower in the POF group than in the control group (0.58 ± 0.38 vs. 1.15 ± 0.67; P < .01). In both groups, there was a significant positive correlation between the mitochondrial DNA/28S rRNA ratio and mitochondrial DNA CT (control group: r = 0.774; P < .001; POF group: r = 0.556; P = .001) and a significant negative correlation between the mitochondrial DNA/28S rRNA ratio and 28S rRNA CT (control group: r = - 0.677; P < .001; POF group: r = - 0.627; P = .001). Conclusion(s): This study has established the clinical feasibility of quantifying amounts of mitochondrial DNA, relative to an internal standard, using real-time PCR. Further studies are warranted to elucidate the roles of apoptosis and mitochondrial function in the pathogenesis of POF.展开更多
Objective: To investigate mutations in the human KIT ligand gene (KITLG) gene as a mechanism of 46,XX spontaneous premature ovarian failure. The human KIT ligand gene, known also as human stem cell factor, is the liga...Objective: To investigate mutations in the human KIT ligand gene (KITLG) gene as a mechanism of 46,XX spontaneous premature ovarian failure. The human KIT ligand gene, known also as human stem cell factor, is the ligand of the c-kit transmembrane tyrosine kinase receptor (KIT). This ligand-receptor interaction is known to play important roles in mouse germ cell migration and proliferation. Design: Cross-sectional study. Setting: Clinical research center. Patient(s): Forty women with 46,XX spontaneous premature ovarian failure. Intervention(s): None. Main Outcome Measure(s): Single-stranded conformational poly-morphism analysis and DNA sequencing. Result(s): We found one nucleotide change of the KITLG coding region (811G→ T) that led to an alteration of the amino acid composition of the KITLG protein in one Caucasian patient (Asp210Tyr). However, we found the same alteration in two normal control Caucasian samples. Three nucleotide substitutions were found in the noncoding exon of KITLG (exon 10). We also identified two intronic polymorphisms. Thus, we did not identify a single significant mutation in the coding region of the KITLG gene in any of 40 patients (upper 95% confidence limit is 7.2% ). Conclusion(s): Mutations in the coding regions of the KITLG gene appear not to be a common cause of 46,XX spontaneous premature ovarian failure in North American women.展开更多
Objective To describe the successful use of preimplantation genetic d iagnosis to assess the prevalence of meiotic errors after oocyte cryopreservation in an o ocyte donation cycle. Design Case report. Setting Private...Objective To describe the successful use of preimplantation genetic d iagnosis to assess the prevalence of meiotic errors after oocyte cryopreservation in an o ocyte donation cycle. Design Case report. Setting Private IVF center. Patient(s) A 42.6-year old patient with ovarian failure. Intervention(s)A donor oocyte IV F cycle with cryopreservation of oocytes followed by thaw, fertilization of oocy tes, preimplantation genetic diagnosis for selective aneuploidy, and ET. Main ou tcome measure(s) Preimplantation genetic analysis of chromosomes 13,16,18, 21,22 , X, and Y with fluorescence in-situ hybridization. Result(s)-The recipients initial serum β-hCG level was 196 mIU/mL 15 days after oocyte retrieval. An i nitial ultrasound at the sixth week of gestation revealed two gestational sacs. A second ultrasound 1 week later showed a monochorionic twin in sac A and a sing leton pregnancy in sac B. Fetal cardiac activity was visualized for all gestatio ns. Conclusion(s) This case illustrates the feasibility of cryopreservation of d onor oocytes combined with preimplantation genetic diagnosis for clinical use in those settings where there may be an increased risk of spindle-related abnorma lities.展开更多
文摘To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group. Design: Prospective study. Setting: Genome Research Center for Reproductive Medicine and Infertility, Korea Ministry of Health & Welfare. Patient(s): Thrity patients with POF and 30 control individuals. Interventions(s): The mitochondrial DNA content was quantified using real-time PCR. The effectiveness of the assay was determined by relative quantification using the comparative threshold cycle (CT) method. Main Outcome Measure(s): Relative quantification of mitochondrial DNA content. Interventions(s): The mitochondrial DNA content was significantly lower in the POF group than in the control group (0.58 ± 0.38 vs. 1.15 ± 0.67; P < .01). In both groups, there was a significant positive correlation between the mitochondrial DNA/28S rRNA ratio and mitochondrial DNA CT (control group: r = 0.774; P < .001; POF group: r = 0.556; P = .001) and a significant negative correlation between the mitochondrial DNA/28S rRNA ratio and 28S rRNA CT (control group: r = - 0.677; P < .001; POF group: r = - 0.627; P = .001). Conclusion(s): This study has established the clinical feasibility of quantifying amounts of mitochondrial DNA, relative to an internal standard, using real-time PCR. Further studies are warranted to elucidate the roles of apoptosis and mitochondrial function in the pathogenesis of POF.
文摘Objective: To investigate mutations in the human KIT ligand gene (KITLG) gene as a mechanism of 46,XX spontaneous premature ovarian failure. The human KIT ligand gene, known also as human stem cell factor, is the ligand of the c-kit transmembrane tyrosine kinase receptor (KIT). This ligand-receptor interaction is known to play important roles in mouse germ cell migration and proliferation. Design: Cross-sectional study. Setting: Clinical research center. Patient(s): Forty women with 46,XX spontaneous premature ovarian failure. Intervention(s): None. Main Outcome Measure(s): Single-stranded conformational poly-morphism analysis and DNA sequencing. Result(s): We found one nucleotide change of the KITLG coding region (811G→ T) that led to an alteration of the amino acid composition of the KITLG protein in one Caucasian patient (Asp210Tyr). However, we found the same alteration in two normal control Caucasian samples. Three nucleotide substitutions were found in the noncoding exon of KITLG (exon 10). We also identified two intronic polymorphisms. Thus, we did not identify a single significant mutation in the coding region of the KITLG gene in any of 40 patients (upper 95% confidence limit is 7.2% ). Conclusion(s): Mutations in the coding regions of the KITLG gene appear not to be a common cause of 46,XX spontaneous premature ovarian failure in North American women.
文摘Objective To describe the successful use of preimplantation genetic d iagnosis to assess the prevalence of meiotic errors after oocyte cryopreservation in an o ocyte donation cycle. Design Case report. Setting Private IVF center. Patient(s) A 42.6-year old patient with ovarian failure. Intervention(s)A donor oocyte IV F cycle with cryopreservation of oocytes followed by thaw, fertilization of oocy tes, preimplantation genetic diagnosis for selective aneuploidy, and ET. Main ou tcome measure(s) Preimplantation genetic analysis of chromosomes 13,16,18, 21,22 , X, and Y with fluorescence in-situ hybridization. Result(s)-The recipients initial serum β-hCG level was 196 mIU/mL 15 days after oocyte retrieval. An i nitial ultrasound at the sixth week of gestation revealed two gestational sacs. A second ultrasound 1 week later showed a monochorionic twin in sac A and a sing leton pregnancy in sac B. Fetal cardiac activity was visualized for all gestatio ns. Conclusion(s) This case illustrates the feasibility of cryopreservation of d onor oocytes combined with preimplantation genetic diagnosis for clinical use in those settings where there may be an increased risk of spindle-related abnorma lities.
