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  • 1
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have extended a previously developed method that allows prenatal DNA diagnosis of female fetuses through the isolation of single nucleated erythrocytes from maternal blood by developing a method that can distinguish between maternal and fetal nucleated erythrocytes. Nucleated erythrocytes were separated by a density-gradient method and then collected by micromanipulation. Sex was determined after primer extension preamplification (PEP) of the entire genome of a single cell, and human leukocyte antigen (HLA)-DQ α type was determined after further amplification of this gene. The HLA-DQ α genotype of fetal erythrocytes in maternal blood samples and their corresponding paternal and maternal lymphocytes were successfully determined in all cases. The accuracy of the method was determined by using single nucleated erythrocytes from umbilical cord blood from five normal deliveries. This is the first demonstration that the fetal HLA-DQ α gene sequences can be identified in a small aliquot of a single nucleated erythrocyte in maternal blood. We believe that this method ushers in a new era in which the reliability and accuracy of noninvasive prenatal DNA diagnosis from maternal blood is markedly improved.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have developed a method that allows the prenatal DNA diagnosis of ornithine transcarbamylase (OTC) deficiency by using a single fetal nucleated erythrocyte (NRBC) isolated from maternal blood. OTC gene analysis of a male patient (TF) with early onset OTC deficiency was performed by single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing. To investigate the possible prenatal diagnosis of OTC deficiency, maternal blood was obtained at 13 weeks of gestation of a subsequent pregnancy, from the mother of patient TF. NRBCs in the maternal blood were separated by using the density gradient method and then collected with a micromanipulator. The entire genome of a single NRBC was amplified by primer extension preamplification (PEP). The human leukocyte antigen (HLA)-DQ alpha genotype and sex were determined from small aliquots of the PEP product. The HLA-DQ alpha genotype of each of the parents of the male patient was also determined. Once a single NRBC had been identified as being of fetal origin, the OTC gene was analyzed by using the restriction fragment length polymorphism (RFLP) method. DNA analysis revealed a point mutation in exon 9 of the OTC gene in the OTC-deficient patient (TF). All NRBCs retrieved from maternal blood were successfully identified as being of fetal origin by HLA-DQ alpha genotyping and sex determination. RFLP analysis demonstrated that the fetal OTC gene was normal. This is the first study to successfully diagnose OTC deficiency prenatally, by using a single fetal NRBC from the maternal circulation. Such prenatal DNA diagnosis is non-invasive and can be applied to other genetic diseases, including autosomal and X-linked diseases.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1436-2813
    Keywords: vaginal delivery ; pelvic floor descent ; incontinence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This study was undertaken to determine the effects of vaginal deliveries on anorectal function, and to analyze the possible clinical, physiological, and radiological risk factors predisposing to damage of the pelvic floor musculature. We studied 25 consecutive women with a mean age of 32 years old, 3 months after vaginal delivery, 17 of whom were primiparae and 8, multiparae. The symptoms of anal incontinence were assessed, and anorectal manometry, rectal sensation, and radiological measurements of the anorectal angle and pelvic floor position at rest, on squeezing, and on straining were performed. As a control, six nulliparous women underwent the same examinations. Pelvic floor descent in both the primiparae and multiparae was significantly greater at rest and on squeezing than that in the nulliparous women. Furthermore, pelvic floor descent on straining was greater in the multiparae than in the nulliparous women (P=0.028). An analysis of the 17 primiparae showed that prolonged duration of the second stage of labor and third-degree perineal tears were important factors predisposing to pelvic floor descent. In fact, 3 of the 17 primiparae (17%) had anal incontinence. These findings indicate that vaginal delivery may cause pelvic floor descent, an obtuse anorectal angle, and bowel symptoms.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1436-2813
    Keywords: Key Words: vaginal delivery ; pelvic floor descent ; incontinence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: P = 0.028). An analysis of the 17 primiparae showed that prolonged duration of the second stage of labor and third-degree perineal tears were important factors predisposing to pelvic floor descent. In fact, 3 of the 17 primiparae (17%) had anal incontinence. These findings indicate that vaginal delivery may cause pelvic floor descent, an obtuse anorectal angle, and bowel symptoms.
    Type of Medium: Electronic Resource
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