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M Adinolfi

Publications and source records attributed to M Adinolfi.

At least 37 records · Page 2Linked to original sources
PubMed1996

Rapid detection of trisomies 21 and 18 and sexing by quantitative fluorescent multiplex PCR.

Aneuploidies involving chromosomes 21, 18, 13, X and Y account for over 95% of all chromosomal abnormalities in live-born infants. Prenatal diagnosis of these disorders is usually accomplished by cytogenetic analysis of amniotic or chorionic cells but this is a lengthy procedure requiring great technical expertise. In this paper, we assess the diagnostic value of using a quantitative fluorescent polymerase chain reaction (PCR) suitable for the simultaneous and rapid diagnosis of trisomies 21 and 18 together with the detection of DNA sequences derived from the X and Y chromosomes. Samples of DNA, extracted from amniotic fluid, fetal blood or tissues, and peripheral blood from normal adults were investigated by quantitative fluorescent PCR amplification of polymorphic small tandem repeats (STRs) specific for two loci on each of chromosomes 21 and 18. Quantitative analysis of the amplification products allowed the diagnosis of trisomies 21 and 18, while sexing was performed simultaneously using PCR amplification of DNA sequences derived from the chromosomes X and Y. These results indicate the advantages of using two sets of STR markers for the detection of chromosome 21 trisomies and confirmed the usefulness of quantitative fluorescent multiplex PCR for the rapid prenatal diagnosis of selected chromosomal abnormalities.

Aneuploidy↗
PubMed1995

Rapid detection of selected aneuploidies by quantitative fluorescent PCR.

Selected aneuploidies can be rapidly diagnosed by the analysis of fluorescent polymerase chain reaction (PCR) products of chromosome-specific and highly polymorphic small tandem repeats (STRs). The quantitative STR patterns obtained from samples of normal individuals are markedly different from those seen when patients with aneuploidies involving chromosome X, or trisomies of chromosomes 21 and 18, are tested. For example, while samples from normal subjects--tested with a chromosome 21-derived STR (D21S11)--show two fluorescent PCR peaks with similar activities in a 1:1 ratio, the analysis of samples from patients with trisomy 21 reveals the presence of either three peaks (ratio 1:1:1), or two peaks with a ratio of 2:1. The use of an internal non-polymorphic marker allows identification of trisomic samples with three copies of the same allele. This rapid approach (24 hours) is particularly valuable when applied to prenatal diagnosis of chromosomal abnormalities since it reduces the time of anxiety of the parents waiting for the results of the conventional cytogenetic tests, which require several weeks.

Aneuploidy↗
PubMed1995

Molecular evidence of fetal-derived chromosome 21 markers (STRs) in transcervical samples.

Transcervical cells (TCCs), collected by flushing or aspiration at 8-13 weeks of gestation, were analysed for the presence of fetal-derived DNA sequences. DNA extracted from maternal peripheral blood, TCC samples, and placental tissue was amplified by the polymerase chain reaction (PCR) to detect small tandem repeat (STR) markers specific to chromosome 21. STR products of fetal origin could be clearly observed in four TCC samples. TCC samples collected by flushing or aspiration were also analysed by fluorescent in situ hybridization (FISH) using X and Y probes simultaneously: 46,XY cells could be detected in all TCC samples obtained from mothers with male fetuses.

Base Sequence↗
PubMed1995

Detection of fetal cells in transcervical samples and prenatal diagnosis of chromosomal abnormalities.

Transcervical samples collected by lavage, aspiration, and cytobrush from women between 6 and 13 weeks of gestation were tested for the presence of fetal cells using fluorescence in situ hybridization (FISH) with probes for chromosomes X, Y, 1, and 21, and by polymerase chain reaction (PCR) amplification of DNA sequences derived from chromosomes X, Y, and 21. With a few exceptions, a good correlation was observed between the results of sexing the fetuses using FISH or PCR on transcervical cell (TCC) samples retrieved by lavage and those obtained by testing fetal (placental) tissue. In a comparative study between TCC samples collected by lavage or cytobrush, the sex of the fetus was correctly diagnosed by PCR amplification of a Y-derived DNA sequence. Variable results were observed with samples obtained by aspiration, mainly because this procedure was found to be more prone to failure to remove thick mucus without previous injection of physiological saline. Chromosome 21-derived small tandem repeats (STRs) of fetal origin were successfully detected in about 40 per cent of TCC samples recovered by lavage. Two cases of chromosomal abnormalities, one of trisomy 21 and one of triploidy, were detected in TCC samples in the course of our investigations.

Cell Separation↗
PubMed1995

Isolation of fetal cells from transcervical samples by micromanipulation: molecular confirmation of their fetal origin and diagnosis of fetal aneuploidy.

Transcervical cell (TCC) samples have been shown to contain fetal cells amenable to molecular analysis. However, the presence of 'contaminating' maternal cells limits their use for prenatal diagnoses. In this report we show that clumps of fetal cells can be isolated from transcervical samples by micromanipulation and tested by fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR). Out of 129 clumps, isolated from mucus aspirates and transcervical lavages from 29 patients, 29 clumps from 11 patients were found to be exclusively of fetal origin as judged by the detection of chromosome 21-specific polymorphic DNA markers and Y-derived DNA sequences by PCR and FISH. One case of a male triploid fetus, diagnosed by the analysis of TCC samples obtained by mucus aspiration and lavage, was confirmed by testing clumps of cells isolated by micromanipulation.

Aneuploidy↗
PubMed1995

Immunohistochemical characterization of cells retrieved by transcervical sampling in early pregnancy.

Trophoblastic cells can be retrieved from the endocervix and the lower uterine segment in early pregnancy by aspiration or lavage (Rodeck et al., 1995). The feasibility of using this technique for prenatal diagnosis depends on how frequently fetal cells can be retrieved and whether such cells can be purified from the predominant maternal cell population. In this study, specimens retrieved from the lower uterine segment prior to elective first-trimester termination of pregnancy were examined histologically and characterized using a panel of monoclonal antibodies in an avidin-biotin-peroxidase technique. Lavage samples generally contained fewer cervical epithelial cells than aspirates. Syncytial fragments or cytotrophoblast were identified in 9 of 12 lavage samples but in only 4 of 10 aspirates. Trophoblast cells were reactive with various anti-trophoblast monoclonal antibodies but the trophoblast cells present displayed considerable antigenic heterogeneity. For positive selection of trophoblast cells from these samples, it is likely that the best yield will be achieved by using a panel of carefully characterized monoclonal antibodies directed against various villous and extravillous trophoblast populations.

Antibodies, Monoclonal↗
PubMed1995

Detection of trophoblast cells in transcervical samples collected by lavage or cytobrush.

OBJECTIVE: To compare two methods of obtaining fetal cells from the endocervical canal for prenatal diagnosis during the first trimester: lavage with physiologic saline, and the endocervical passage of a cytobrush. METHODS: Fetal cells were identified morphologically using conventional and immunohistochemical staining. Y-specific sequences were detected using polymerase chain reaction (PCR) and fluorescent in situ hybridization. The presence of fetal DNA was also demonstrated by PCR amplification of a polymorphic short tandem repeat marker. RESULTS: Syncytiotrophoblast cells were identified in four of 11 lavage samples and in one of 11 brush samples. Sex determination using fluorescent in situ hybridization was achieved in all 11 lavage samples and in nine of 11 cytobrush samples; no Y sequences were located in two brush samples from male embryos. Sex determination was achieved successfully in all 22 samples using PCR of X- and Y-specific DNA sequences. Small tandem repeat pattern analysis indicated the presence of fetal DNA in five of 11 lavage samples and in one of eight cytobrush samples, which were informative because the maternal and fetal small tandem repeat patterns differed. CONCLUSION: Although lavage retrieves more trophoblast cells than does brushing, both methods are potentially suitable for molecular prenatal diagnosis.

Female↗
PubMed1994

Composition of the coagulant polysaccharide fraction from Strychnos potatorum seeds.

The composition of the coagulant polysaccharide fraction from Strychnos potatorum seeds is described. This fraction comprises a 1:1.7 mixture of a galactomannan and a galactan. The structure of these polysaccharides is also discussed. In addition, the coagulant properties of the polysaccharide fractions of two other Strychnos species, innocua and nux-vomica, have been assayed.

Carbohydrate Conformation↗
PubMed1994

Rapid molecular method for prenatal detection of Down's syndrome.

We have evaluated a rapid method that allows prenatal detection of Down's syndrome in less than 24 hours. DNA from uncultured amniotic fluid, fetal blood, and tissue samples was amplified with the small tandem repeat (STR) marker D21S11. Quantitative analysis of fluorescent STR products with evaluation of their sizes provided clear evidence for trisomy 21. Whilst most normal samples showed two amplification peaks of equal size, Down's syndrome samples were characterised by either three STR peaks or two peaks with a ratio of 2:1. Co-amplification with a non-polymorphic sequence allowed analysis of samples that were homozygous for the 21-derived STRs.

Amniotic Fluid↗
PubMed1994

FISH detection of trisomy 21 in interphase by the simultaneous use of two differentially labelled cosmid contigs.

Techniques have been reported in which fluorescence in situ hybridisation (FISH) and cosmid probes are used to detect trisomy 21 (and other abnormalities involving chromosomes X, Y, 13, and 18) on uncultured amniocytes. However the detection rate of trisomy 21 is lower than for the other anomalies owing to a larger number of uninformative results and false negatives. We report the simultaneous use of two differentially labelled cosmid contigs to improve the detection rate of trisomy 21 on uncultured amniocyte samples thus allowing the prenatal diagnosis of Down's syndrome even if only few labelled nuclei are available.

Amniotic Fluid↗
PubMed1993

Detection of trisomy 18 and Y-derived sequences in fetal nucleated cells obtained by transcervical flushing.

A procedure which combines the collection of fetal cells by transcervical flushing and in situ hybridisation techniques on nuclei in interphase was used to detect trisomy 18 in a fetus at 12 weeks of gestation. Using a primed in situ labelling method, we could also detect Y-specific sequences in a small percentage of transcervically flushed cells obtained at 8-12 weeks from pregnant women with male fetuses. This approach seems to be suitable for prenatal diagnosis of major chromosomal abnormalities and other selected inherited disorders very early in gestation.

Adult↗
PubMed1993

Triterpenoid oligoglycosides from Chionodoxa luciliae.

Four minor novel oligoglycosides of triterpenes having a lanostane-type skeleton were isolated from the bulbs of Chionodoxa luciliae. The C30 skeleton of the aglycones has been found for the first time in Liliaceae and can be considered the metabolic precursor of the C29 eucosterol skeleton.

Carbohydrate Sequence↗