bluuebunny/crossref_metadata_2025 huggingface dataset:
Alpha‐fetoprotein and acetylcholinesterase activity in first‐and early second‐trimester amniotic fluid
Authors: Stener Jørgensen Finn , Sundberg Karin , Loft Anne Gitte Rasmussen , Arends Jørgen , Nørgaard‐Pedersen Bent
Published: March 2007

Normal ranges of amniotic fluid alpha‐fetoprotein (AFP) and acetylcholinesterase activity (AChE) are described for gestational weeks 11–14 using rocket gel immunoelectrophoresis for AFP quantitation and a monoclonal antibody (4F19) enzyme antigen immunoassay for AChE activity measurement. The normal ranges were established by the examination of 281 amniotic fluid samples from 281 normal pregnancies. AFP was found to increase from a median level of 14.0 MIU/1 at 11 weeks to a maximum at 13 weeks (median=18.0 MIU/l) (P<0.05), thereafter falling (not significant). No AChE test result exceeded 4.8 nkat/l. In addition, AFP and AChE values for three cases of fetal malformation, identified by the biochemical analyses of amniotic fluid, are given. These cases included two fetuses with a neural tube defect and one fetus with an abdominal wall defect. Amniocentesis was performed at 10, 11, and 14 weeks, respectively. The AFP and AChE values were all high.
The significance of early second‐trimester sonographic detection of minor fetal renal anomalies
Authors: Bronshtein Moshe , Bar‐Hava Itai , Lightman Abraham
Published: March 2007

A study of 6350 consecutive transvaginal ultrasound examinations was performed as part of a routine fetal evaluation. Twenty‐one cases (0.33 per cent) of early second‐trimester sonographic detection of minor renal abnormalities (unilateral renal agenesis, pelvic kidney, and double collecting system) are presented. The sonographic diagnosis was made at 14–18 weeks of pregnancy and confirmed, in all of the 21 fetuses, postnatally or by post‐mortem. A high incidence of associated fetal anomalies (24 per cent) and parental renal abnormalities (14 per cent) was demonstrated. Transvaginal sonography was found to be a useful tool for diagnosing these renal anomalies as early as 14 weeks of pregnancy. The likelihood of various associated anomalies and long‐term implications on renal function raise questions concerning the prenatal management of such patients.
Prenatal findings in generalized amyoplasia
Authors: Sepulveda Waldo , Stagiannis Konstantinos D. , Cox Phillip M. , Wigglesworth Jonathan S. , Fisk Nicholas M.
Published: March 2007

Amyoplasia is a rare, sporadic condition characterized by different degrees of maldevelopment of the skeletal muscles, which are replaced by fibrous and fatty tissue. In this report, we present a case of generalized amyoplasia presenting at 19 weeks' gestation. The most striking finding was the absence of fetal movements, resulting in severe multiple congenital contractures, hydrops, and polyhydramnios. At autopsy, histological examination of the skeletal muscle showed small groups of poorly developed fibres within areas of fat. This report suggests that generalized amyoplasia could be a common cause of severe forms of multiple congenital contractures, but is probably underdiagnosed at post‐mortem because of inadequate examination of muscles. Definitive diagnosis is important in determining the risks of recurrence in these cases.
Prenatal analysis of the insulin receptor gene in a family with leprechaunism
Authors: Krook A. , Bell J. A. , Robertson M. E. , Brueton L. , O'Rahilly S.
Published: March 2007

We report on the prenatal diagnosis of a fetus at risk of leprechaunism. We had previously determined the nature of the causative mutation in the insulin receptor gene in this family. The mutation removes a restriction site for the enzyme Mbo II. Genomic DNA was extracted from a chorionic villus sample and the 3′ half of exon 2 was amplified by the polymerase chain reaction (PCR) followed by restriction digest. Using this method, we correctly predicted an unaffected child.
Carrier detection of duchenne muscular dystrophy through analysis of dna from deciduous teeth of a dead affected child
Authors: Restagno Gabriella , Ferrone Marina , Doriguzzi Carlo , Palmucci Laura , Mongini Tiziana , Carbonara Angelo
Published: March 2007

The sister of a child affected by Duchenne muscular dystrophy (DMD) was referred for genetic counselling to assess the risk of her being a carrier. Her brother had died 15 years previously at the age of 8. There were no other affected males in the family. There were no methods for DNA investigation at the time of the child's death and the family had never been studied for linkage with polymorphic probes on the chromosomal region Xp21. The only tissue from which an assessment of the risk could be made by DNA linkage analysis was two of the child's deciduous teeth that the parents had kept. DNA was extracted using a protocol described for the recovery of ancient DNA from museum specimens and archaeological finds. Multiplex amplification did not reveal deletions in 19 exons spanning the hot‐spot regions for deletions within the dystrophin gene in Xp21. Linkage analysis using three highly polymorphic microsatellites demonstrated that the sister had not received the X chromosome borne by her brother. These results show that DNA extracted from teeth is a reliable source for molecular diagnosis.
Crown—rump length in aneuploid fetuses: Implications for first‐trimester biochemical screening for aneuploidies
Authors: Macintosh M. C. M. , Brambati B. , Chard T. , Grudzinskas J. G.
Published: March 2007

This study examined the effect of estimation of gestational age from the menstrual history compared with that from crown—rump length (CRL) measurement on the detection rate of screening for aneuploidies in the first trimester. Pregnancy‐associated plasma protein A (PAPP‐A) was assayed in blood collected prior to chorionic villus sampling in 356 women with unaffected pregnancies and 28 women with an aneuploid pregnancy. There were 14 Down's syndrome (DS) pregnancies. All pregnancies were dated from menstrual history and CRL measurement. The average CRL gestation in the aneuploid population was 2.5 days less than that derived from the LMP (95 per cent confidence interval (CI) for LMP—CRL gestation: using the algorithm based on unaffected pregnancies 0–3.5 days; using the matched case—control approach 1–4.5 days). The average CRL gestation in the DS population was 2 days less but this did not reach statistical significance (95 per cent CI for LMP—CRL gestation: using the algorithm — 1 to 4.5 days; using the matched case—control approach 0 to 5.5 days). The detection rate of aneuploidies in the first trimester using maternal serum PAPP‐A was reduced by 7 per cent (and by 3 per cent for DS) for a 5 per cent false‐positive rate when using CRL rather than LMP to date the pregnancy. This phenomenon is a consequence of an apparent reduction of gestational age when estimated by CRL in the aneuploid population. Further studies are required to evaluate whether CRL is an unbiased estimate of gestation for Down's syndrome pregnancies.
A chromosome 21‐specific cosmid cocktail for the detection of chromosome 21 aberrations in interphase nuclei
Authors: Van Opstal Diane , Van Hemel Jan O. , Eussen Bert H. J. , Van Der Heide Annet , Van Den Berg Cardi , Veld Peter A. In't , Los Frans J.
Published: March 2007

Fluorescent in situ hybridization (FISH) with a 21q11‐specific probe (CB21c1) consisting of three non‐overlapping cosmids has been applied to interphase amniocytes of pregnancies at increased risk for fetal aneuploidy (N = 78) and to interphase lymphocytes, cultured and uncultured, of patients referred for Down syndrome (N = 19 and 28, respectively). In the uncultured amniocytes, six chromosome aberrations were detected: three cases of trisomy 21, a triploidy, a de novo 46,XX,t(21q21q), and a mosaic 46,XY/47,XY,+dic(21)(q11)/48,XY,+dic(21)(q11), +del(21)(q11). In 15 cultured and 20 uncultured blood samples, FISH correctly diagnosed trisomy 21 (full or mosaic) at the interphase level, which was confirmed in all cases by subsequent karyotyping. Because of specific and strong signals in interphase nuclei, CB21c1 appears to be a useful tool for the rapid detection of chromosome 21 abnormalities.
Molecular prenatal diagnosis of 3‐hydroxy−3‐methylglutaryl coa lyase deficiency
Authors: Mitchell Grant A. , Jakobs Cornelis , Gebson K. Michael , Robert Marie‐France , Burlina Alberto , Dionisi‐Vici Carlo , Dallaire Louis
Published: March 2007

We report the first molecular prenatal diagnosis of 3‐hydroxy‐3‐methylglutaryl CoA lyase (HL) deficiency. The proband had a classic but severe presentation with hypoketotic hypoglycaemia and acidosis, secondary mental retardation, and epilepsy, and HL deficiency was documented in cultured fibroblasts. We found him to be homozygous for the frameshift mutation N46fs (+1), which yields a distinct pattern on single‐strand conformation polymorphism (SSCP) analysis. In two subsequent pregnancies, molecular prenatal diagnosis was performed using SSCP. In the first, chorionic villus biopsy was normal. In the second pregnancy, amniocentesis revealed an affected fetus. In both pregnancies, the diagnosis was confirmed enzymatically. HL activity was less than 7 per cent of control values in amniocytes and fetal liver of the affected pregnancy. In the second pregnancy, amniotic fluid metabolite measurements by stable isotope dilution‐selected ion monitoring mass spectrometry showed greater than 100‐fold increases of 3‐hydroxy‐3‐methylglutaric acid and of 3‐methylglutaconic acid levels compared with controls.
Second‐trimester levels of maternal urinary gonadotropin peptide in down syndrome pregnancy
Authors: Canick Jacob A. , Kellner Leonard H. , Saller Devereux N. , Palomaki Glenn E. , Walker Roger P. , Osathanondh Rapin
Published: March 2007

Urinary gonadotropin peptide (UGP; β‐core fragment), a major metabolite of human chorionic gonadotropin (hCG), was shown recently to be markedly elevated in Down syndrome pregnancy between 19 and 22 weeks of gestation. To confirm and extend this finding, we obtained maternal urine and matching maternal serum samples from 14 cases of Down syndrome and six other aneuploidies between 17 and 21 weeks of gestation. UGP was measured in all these samples and in 91 singleton control urines. Results were corrected for urinary creatinine level and expressed as multiples of the control median (MOM). hCG levels were assayed in all serum samples from the cases and compared with previously established reference values. The median UGP level in Down syndrome cases was 5.34 MOM (range 2.71–12.57); 88 per cent of the values were above the 95th centile of control levels after modelling. The median maternal serum hCG level for the same cases was 2.20 MOM (range 0.84–3.40); 36 per cent of the values were above the 95th centile. The level of UGP in every case including all other aneuploidies was higher than the comparable maternal serum hCG level. Elevated UGP measurements are strongly associated with fetal Down syndrome during the second trimester and could contribute to improved Down syndrome screening protocols that are more accessible and less expensive than are currently available.
Urinary multiple marker screening for down's syndrome
Authors: Cuckle H. S. , Iles R. K. , Sehmi I. K. , Chard T. , Oakey R. E. , Davies S. , Ind T.
Published: March 2007

We have examined the possibility of using multiple markers in maternal urine rather than serum in order to screen for Down's syndrome. Urine samples were available from 36 cases (24 Down's syndrome, five Edwards' syndrome, three Turner's syndrome, one Klinefelter's syndrome, one triploidy, one triple‐X, one twin discordant for Down's syndrome) and 294 controls, including three twins. Three markers were tested: the β‐core fragment of human chorionic gonadotrophin (hCG), total oestrogen (tE) and the free a subunit of hCG. Levels were corrected for creatinine excretion and expressed as multiples of the gestation‐specific median (MOM) level from the singleton controls. The median value for the singleton Down's syndrome cases was 6.02, 0.74, and 1.08 MOM for β‐core‐hCG, tE, and a‐hCG, respectively. The increases in β‐core‐hCG and the reduction in tE levels were highly significant (P<0.0001 and 0.005, respectively; Wilcoxon rank sum test) but the increase in free a‐hCG was not (P=0.40). On the basis of a mathematical model, the expected detection rate for a 5 per cent false‐positive rate was 79.6 per cent for β‐core‐hCG alone, which increased to 82.3 per cent when combined with tE. Aneuploidies other than Down's syndrome were characterized by low levels of tE and either low or high β‐core‐hCG.
Prenatal diagnosis of metatropic dwarfism
Authors: Manouvrier‐Hanu S. , Devisme L. , Zelasko M. C. , Bourgeot Ph. , Vincent‐Delorme C. , Valat‐Rigot A. S. , Puech F. , Farriaux J. P.
Published: March 2007

We present a case of prenatal diagnosis of severe metatropic dysplasia at 20 weeks' gestation. The characteristic prenatal features of this rare autosomal recessive chondrodysplasia appear to be significant dwarfism with an enlarged head and a narrow thorax associated with enlargement of the hands and feet, and the radiographic ‘dumb‐bell’ appearance of the long bones.
Apparent segregation of null alleles ascribed to deletions of the ornithine transcarbamylase gene in congenital hyperammonaemia
Authors: Segues Bertrand , Rozet Jean‐Michel , Gilbert Brigitte , Saugier‐Veber Pascale , Rabier Daniel , Saudubray Jean‐Marie , Carré Mireille , Rouleau Françoise Parrot , Menget Alain , Bonardi Jean‐Michel , Lyonnet Stanislas , Bonnefont Jean‐Paul , Munnich Arnold
Published: March 2007

Ornithine transcarbamylase (OTC) deficiency is an X‐linked disorder of the urea cycle mapped to chromosome Xp21.1. Here, we show that apparent segregation of null alleles at the OTC locus and flanking polymorphic loci mimicked false maternity or false paternity in three affected families. Based on these observations, we suggest giving consideration to gene deletion when dealing with segregation of null alleles in OTC deficiency.
Ultrasound diagnosis of the pena shokeir phenotype at 14 weeks of pregnancy
Authors: Ajayi R. A. , Keen C. E. , Knott P. D.
Published: March 2007

This report describes the early prenatal diagnosis of the Pena Shokeir phenotype in an at‐risk patient at 14 weeks' gestation. The diagnosis was based on an abnormal fetal movement profile, in association with an abnormal position of the fetal limbs. Pena Shokeir phenotype describes an inherited condition characterized by arthrogryposis and dysmorphic features as a result of fetal akinesia. It is a lethal abnormality and early diagnosis allows safer surgical methods of termination.
Prenatal diagnosis of trisomy 8 mosaicism in cvs after abnormal ultrasound findings at 12 weeks
Authors: Guichet A. , Briault S. , Toutain A. , Paillet C. , Descamps P. , Pierre F. , Body G. , Moraine Cl.
Published: March 2007

We describe a case of trisomy 8 mosaicism in which fetal chromosome analysis was prompted by ultrasound abnormalities, i.e., hygroma colli and dilatation of the renal pelves. Chorionic villus sampling (CVS) was performed, with a false‐negative result on direct karyotype analysis, although cultured trophoblasts revealed trisomy 8 mosaicism. Fetal autopsy confirmed the abnormalities found on ultrasound examinations and fetal tissue examination showed different levels of trisomy 8 mosaicism. To our knowledge, this is the first prenatal diagnosis of trisomy 8 made on ultrasound findings.
Prenatal diagnosis of fragile x syndrome: (cgg)n expansion and methylation of chorionic villus samples
Authors: Castellví‐Bel Sergi , Milà Montserrat , Soler Anna , Carrió Ana , Sánchez Aurora , Villa Margarita , Jiménez M Dolores , Estivill Xavier
Published: March 2007

Fragile X syndrome is the most common form of inherited mental retardation, due to an expansion of the (CGG)n trinucleotide repeat in the FMR‐1 gene and hypermethylation of its 5′ upstream CpG island. Two major problems remain to be resolved for fragile X prenatal diagnosis: the abnormal methylation patterns of chorionic villus samples (CVS) and the inability to predict the mental status of females with the full mutation. We present here the results of ten prenatal diagnoses of fragile X syndrome using Southern blotting and polymerase chain reaction (PCR) amplification, and the analysis of 50 further CVS to test the methylation status of the CpG island of the FMR‐1 gene. In the ten ‘at‐risk’ CVS, eight normal (five males and three females) and two affected male fetuses were detected. Absence of methylation in the CVS was observed in two cases, which was not found upon subsequent examination of the newborn or of fetal tissues. In the 50 CVS not ‘at risk’ for fragile X syndrome, abnormal fragment patterns for probe StB12.3 were detected in 32 per cent for female and 24 per cent for male fetuses. This abnormal pattern could be due to absent or partial methylation of the CpG island of the FMR‐1 gene in chorionic villus tissues.
Pregnancy outcome after transcervical cvs with a flexible biopsy forceps: Evaluation of risk factors
Authors: Lunshof Simone , Boer Kees , Leschot Nico J. , Pomp Marc , Wolf Hans
Published: March 2007

The pregnancy outcome of 1936 women who had transcervical chorionic villus sampling (CVS) with a flexible biopsy forceps was evaluated. Follow‐up until 4 weeks after delivery was 99.4 per cent. Various patient‐ and procedure‐related risk factors for spontaneous loss (fetal or neonatal death) were analysed using stepwise logistic regression analysis. The overall spontaneous loss rate was 4.5 per cent. Factors found to be significantly associated with spontaneous loss were quantity of villi ≤ 15 mg (relative risk (RR) 2.13), a history of first‐trimester miscarriage (RR 1.87) or delivery between 16 and 27 weeks (RR 3.87), cervical culture positive for anaerobes (RR 4.52) or group B streptococcus (RR 3.62), post‐procedural bleeding >3 days (RR 1.99), and multiple insertions (RR 2.64). Significant differences in loss rates between individual operators were found. A learning effect was not present. There were no infants born with terminal transversal limb anomalies in our series.
Isolated fetal hydronephrosis: Beware the effect of bladder filling
Authors: Petrikovsky Boris M. , Cuomo Margaret I. , Schneider Elizabeth P. , Wyse Linda J. , Cohen Harris L. , Lesser Martin
Published: March 2007

The aim was to assess the role that fetal bladder size has in the determination of fetal hydronephrosis. Forty‐three fetuses were evaluated for fetal hydronephrosis in the second trimester of pregnancy. Anteroposterior measurements of the renal pelvis were obtained with a full bladder and again when the bladder emptied in each fetus. Statistical analysis was performed using the Spearman rank order correlation coefficient to assess the relationship between bladder status and renal dilation. The anteroposterior size of the fetal renal pelvis diminished from 6.8 ± 1.8 mm on a full bladder scan to 4.5 ± 1.6 mm when the bladder was emptied (P<0.001). Fifty‐three per cent of the fetuses whose renal pelvic measurements were 5 mm or more on a full bladder scan had normal‐appearing renal pelvises when their bladders emptied. The status of the fetal bladder should be considered when evaluating fetal hydronephrosis.
Chromosome 18 analysis by fluorescence in situ hybridization (fish) in human blastomeres of abnormal embryos after in vitro fertilization (ivf) attempt
Authors: Bergere M. , Selva J. , Baud M. , Volante M. , Martin B. , Hugues J. N. , Olivennes F. , Frydman R. , Auroux M.
Published: March 2007

We performed fluorescence in situ hybridization (FISH) with a chromosome 18‐specific probe on human abnormal cleaved embryos, fertilized either by two spermatozoa and exhibiting three pronuclei (3 PN) or normally fertilized and exhibiting two pronuclei (2 PN) with subsequent severe fragmentation and/or blocking. The aim of the study was to evaluate the incidence of chromosome 18 anomalies among these embryos, in order to evaluate the FISH efficiency on such material and to obtain more precise and complete data than those obtained with classical cytogenetic analysis. For the 3 PN cleaved embryos, FISH confirmed the frequent regulation towards diploidy (25 per cent) and the high frequency of mosaics (53 per cent). For the 2 PN blocked or damaged embryos, FISH permitted chromosome evaluation, which was otherwise impossible with classical cytogenetic techniques: we also found a high mosaic frequency (45 per cent) with these embryos. If this frequency were the same for normally developing embryos, it would be a major obstacle to the reliability of either chromosomal or genetic preimplantation diagnosis.
The prognostic factors in the prenatal diagnosis of the echogenic fetal lung
Authors: Barret Jon , Chitayat David , Sermer Matthew , Amankwah Koffi , Morrow Robert , Toi Ants , Ryan Greg
Published: March 2007

The prenatal diagnosis of an echogenic fetal lung (EFL) is now often made in the early second trimester using high‐resolution ultrasound. This ultrasound appearance is usually caused by a congenital cystic adenomatoid lung malformation (CCAM), an intrapulmonary lung sequestration or obstruction of a major airway. In order to provide prognostic guidelines to parents who may be considering termination of a fetus with these findings, we have analysed a series of 11 cases diagnosed in our centre over the past 2 years in conjunction with 60 cases from major published series. The data suggest that in the absence of non‐immune hydrops fetalis (NIHF) or other anomalies, the outcome for the fetuses is excellent, with over 90 per cent survival. Neither early diagnosis (24 weeks) nor the presence of mediastinal shift is a poor prognostic indicator. In addition, it appears that if NIHF is absent at diagnosis, the chance that it will develop as the pregnancy continues is small (6 per cent). Furthermore, there is a significant (up to 30 per cent) chance that this ultrasound finding will resolve in utero. The development of in utero fetal surgical techniques may be the only hope for those hydropic fetuses who appear to have a dismal prognosis.
Prenatal diagnosis of smith–lemli–opitz syndrome is possible by measurement of 7‐dehydrocholesterol in amniotic fluid
Authors: Dallaire L. , Mitchell G. , Giguère R. , Lefebvre F. , Melançon S. B. , Lambert M.
Published: March 2007

Amniocentesis was performed at 17.3 weeks in a pregnancy with severe intrauterine growth retardation. Cytogenetic studies on amniocytes were normal, 46,XX, and the pregnancy was continued. The diagnosis of Smith–Lemli–Opitz syndrome was suspected in the neonatal period and confirmed by the presence of 7‐dehydrocholesterol (7‐DHC) in the plasma (0.4 mmol/l, normal = not detectable) associated with a low total cholesterol concentration (0.4 mmol/l, normal = 2.56 ± 0.23). Retrospective analysis of the amniotic fluid sample revealed an elevated level of 7‐DHC (0.022 mmol/l; normal = undetectable). Therefore measurement of 7‐DHC levels in amniotic fluid during the second trimester of pregnancy is useful for the prenatal diagnosis of Smith–Lemli–Opitz syndrome in families at risk and should be considered in cases of severe growth retardation of unknown aetiology for which amniotic fluid is available and in which a normal chromosomal pattern in amniocytes is present.
Flow sorting of fetal erythroblasts using intracytoplasmic anti‐fetal haemoglobin: Preliminary observations on maternal samples
Authors: Zheng Yun‐Ling , Demaria Maryann , Zhen Dongkai , Vadnais Theresa J. , Bianchi Diana W.
Published: March 2007

Monoclonal antibody to fetal haemoglobin (a2γy2) has been proposed as a fetal‐specific reagent. We developed an intracellular staining protocol that combines fluorescein isothiocyanate or phycoerythrin conjugated anti‐γ with the DNA binding dye Hoechst 33342 to identify and flow sort fetal erythroblasts from maternal blood. Our preliminary observations on anti‐γ‐positive cells sorted from four different pregnant women are described here, using fluorescence in situ hybridization (FISH) with chromosome‐specific probes to identify fetal cells. Our data demonstrate that far fewer candidate fetal cells are sorted with this protocol than by current cell surface staining methods that employ the monoclonal antibody CD71. This results in increased fetal cell sorting purities. With this protocol, standard FISH techniques require modification due to the rigorous fixation with 4 per cent paraformaldehyde. Our initial data indicate the promise of this approach.
Isolating fetal nucleated red blood cells from maternal blood: The baylor experience—1995
Authors: Simpson Joe Leigh , Lewis Dorothy E. , Bischoff Farideh Z. , Elias Sherman
Published: March 2007

In our previous work we have isolated fetal cells from maternal blood and used fluorescent in situ hybridization (FISH) for chromosome‐specific probes to detect aneuploidy. Current efforts in the Baylor College of Medicine programme are focusing on obtaining consistency in flow‐sorting methodology and on determining sensitivity and specificity. To this end, systematic evaluation of five glycophorin A (gly A) antibodies all produced agglutination, leading us to abandon the use of gly A antibodies for positive selection of fetal cells. Conversely, we have found LDS‐751 to be useful for nuclear selection. CD45 negative selection can best be accomplished by the use of flasks coated with goat antibodies against mouse antibodies. Positive selection by flow sorting for either CD71+ cells or gamma‐globin‐positive cells seems to be successful. Using these two approaches, we have recently detected male (fetal) cells in pregnancies in which the fetus was 46, XY in 10 of 18 and in 12 of 14 cases, respectively.
Enrichment of fetal nucleated cells from maternal blood: Model test system using cord blood
Authors: Andrews Kathryn , Wienberg Johannes , Ferguson‐Smith Malcolm A. , Rubinsztein David C.
Published: March 2007

The presence of small numbers of fetal nucleated red cells in the maternal circulation has been a stimulus for the development of technologies for non‐invasive prenatal genetic analysis. Our laboratory has been assessing the feasibility of density gradient centrifugation followed by magnetic activated cell sorting (MACS) of cells expressing CD32 and CD45, to deplete maternal nucleated blood cells. We have examined the efficiency of each of the steps of this procedure using cord blood from term pregnancies as a source of nucleated red blood cells. Cord blood was shown to contain highly variable numbers of nucleated red cells. Three different density gradients were examined. There was no major difference in the performances of the double and triple gradients. Density gradient centrifugation resulted in enrichments of nucleated red blood cells of about 1000‐fold relative to the total cell count. However, it was apparent that the selection of the cell layers which were most enriched for these cells would result in significant losses of nucleated red cells in other layers. MACS sorting of cells using CD45 resulted in white cell depletions ranging from 7 to 34‐fold. These data provide a foundation for comparison with other methods and for optimization of the MACS technique.
Methods for the transcervical collection of fetal cells during the first trimester of pregnancy
Authors: Rodeck Charles , Tutschek Boris , Sherlock Jon , Kingdom John
Published: March 2007

Before termination of pregnancy, four techniques for retrieving fetal cells transcervically were investigated: uterine lavage, endocervical lavage, cytobrush and mucus aspiration. The yield of fetal cells in these samples was studied and found to be somewhat better after uterine lavage. A preliminary assessment of the safety of mucus aspiration was carried out before transcervical chorionic villus sampling (CVS) in continuing pregnancies. No difference in outcome to a control group having only CVS was found.
Amniocentesis rate and the detection of down syndrome and other chromosomal anomalies in Israel
Authors: Shohat Mordechai , Akstein Edna , Davidov Bella , Barkai Gad , Legum Cyril , David Miriam , Dar Hanna , Romem Yitzhak , Amiel Aliza , Cohen Hana , Bach Gideon , Ben‐Neriah Ziva , Sheffer Ruth N. , Appelman Zvi , Chemke Juan , Zadka Pnina , Zer Tamy , Goldman Boleslaw
Published: March 2007

We investigated the contribution of different screening criteria to the prenatal detection of Down syndrome (DS) as well as other chromosomal anomalies in the Jewish population in Israel during 1990 and 1992. There was a significant decrease (P<0·03) in the incidence of DS live‐births during 1992 (40:78 442) compared with 1990 (69:73 751) which paralleled a marked increase in total prenatal testing and in DS cases detected prenatally. Private laboratories, which perform amniocenteses mostly for women with a low risk of DS and without genetic counselling, had a significantly lower detection rate (1:917) compared with that of the genetic institutes, which following genetic counselling test both women ≥ 37 years of age (1:91) and women younger than 37 years (1:113). The detection of chromosomal anomalies other than DS was less affected by the reason for amniocentesis. Amniocentesis indicated by maternal serum marker screening of women younger than 37 years identified a greater number of chromosomal anomalies other than DS than amniocentesis based on age (≥37 years) alone (111:9604 versus 94:9810; P<0·06). Prenatal detection of DS is most effective when the indication for amniocentesis follows genetic counselling. The increasing use of maternal serum marker screening leads to a significant improvement in the positive detection rate of chromosomal anomalies other than DS in young women.
Early amniocentesis and fetal nuchal translucency in women requesting karyotyping for advanced maternal age
Authors: Salvesen Douglas R. , Goble Orla
Published: March 2007

Fetal nuchal translucency was measured at 11–14 weeks' gestation in 97 pregnancies referred for early amniocentesis for advanced maternal age. The nuchal translucency was abnormal in 11 fetuses and the fetal karyotype was abnormal in five of these 11 cases. The karyotype was normal in 86 cases with normal nuchal translucency. The culture failure and miscarriage rates associated with early amniocentesis were 3·3 per cent and 2·2 per cent respectively. Amniotic fluid leakage occurred in 6 per cent of cases. In women requesting fetal karyotyping for advanced maternal age without additional biochemical screening, fetal nuchal translucency should be measured at 11–14 weeks. If the nuchal thickness is ≥ 3 mm, a first‐trimester diagnostic procedure is indicated; however, if it is <3 mm, amniocentesis should be delayed until 16 weeks' gestation.
Complications of cordocentesis in high‐risk pregnancies: Effects on fetal loss or preterm delivery
Authors: Bernaschek Gerhard , Yildiz Akgun , Kolankaya Aytug , Stuempflen Ingrid , Deutinger Josef
Published: March 2007

Between 1990 and 1993, 166 cases underwent cordocentesis and were followed for at least the following 4 weeks in the Prenatal Diagnosis and Therapy Centre of Vienna University. The indications for the procedure were structural malformations in 46·4 per cent of the cases, other high‐risk diagnoses in 48·8 per cent, and maternal age over 35 years in only 4·8 per cent. We investigated retrospectively all cases of complications resulting in fetal loss or preterm labour. Abortion, intrauterine fetal death, chorioamnionitis, and preterm delivery occurred in 0·6, 5·4, 0·6 and 9·0 per cent of these cases, respectively, adding up to a total of 26 cases (15·7 per cent). Although this rate looks relatively high, 20 of the 26 cases had already displayed signs implying a complicated prognosis. Neither maternal age, gestational age, number of attempts, nor placental location correlated with fetal loss or preterm delivery. Significantly higher rates of fetal loss or preterm delivery were observed when cordocentesis was performed in cases diagnosed as duodenal/intestinal stenosis or hydrops–ascites–hydrothroax/hygroma colli (P=0·0488 and P=0·0005). The frequency of complications did not decrease as the experience of the operators increased.
Cardiac flow after fetal blood sampling in normally grown and growth‐retarded fetuses
Authors: Capponi Alessandra , Rizzo Giuseppe , Rinaldo Donatella , Arduini Domenico , Romanini Carlo
Published: March 2007

The objective of this study was to evaluate the effect of fetal blood sampling on cardiac flow velocity waveforms. Flow velocity waveforms were measured from the ascending aorta and pulmonary artery immediately before and after fetal blood sampling in 29 normally grown and 12 growth‐retarded fetuses. The latter group was characterized by abnormal Doppler indices in the umbilical artery and middle cerebral artery suggestive of uteroplacental insufficiency as the causative factor of the impaired growth. The flow velocity parameters studied were the peak velocity, the time to peak velocity, and the left and right cardiac output and their ratio. In normally grown fetuses, the peak velocity and right and left cardiac output values increased significantly after fetal blood sampling, while no significant changes were observed in the other indices considered. The gestational age at the time of the procedure was positively related to the amplitude of these changes. In growth‐retarded fetuses, fetal blood sampling did not induce any significant increase in cardiac output or peak velocities, while in more than 50 per cent of the fetuses these Doppler indices decreased. The amplitude of the decrease was significantly related to the severity of acidosis in the umbilical vein. In conclusion, the cardiac haemodynamic response to fetal blood sampling differs between normally grown and growth‐retarded fetuses. This difference may explain the higher rate of complications occurring in the latter group of fetuses after blood sampling.
Mosaicism for trisomy 12: Four cases with varying outcomes
Authors: Bischoff Farideh Z. , Zenger‐Hain Julie , Moses David , van Dyke Daniel L. , Shaffer Lisa G.
Published: March 2007

Trisomy 12 observed in chorionic villus sampling (CVS) may reflect generalized mosaicism or indicate mosaicism confined to only the placenta. In this report, four cases of trisomy 12 observed in CVS or cultured placental biopsies with varying outcomes are presented. Seven dinucleotide repeat polymorphisms for chromosome 12 were used to determine the chromosome 12 origins in the fetus or child and to delineate the mechanism(s) that gave rise to the trisomy. In two cases (cases A and C), the mosaicism was confined to the placenta, resulting in normal liveborns. Although, in one case, the molecular results suggested an apparent duplication of one paternal chromosome 12 in the placenta, normal biparental inheritance was found in the diploid fetal cell line in both cases. In two other cases (cases B and D), trisomy 12 was observed in both extraembryonic and fetal tissues. In one of these pregnancies, a child was born by Caesarean section at 37 weeks because of intrauterine growth retardation and oligohydramnios, and resulted in neonatal death. Molecular markers and fluorescence in situ hybridization (FISH) revealed low‐level trisomy 12 mosaicism in the spleen. In the fourth case, fetal abnormalities were detected on ultrasound and low‐level trisomy 12 mosaicism was observed in amniotic fluid cells using conventional cytogenetics and FISH. Molecular markers revealed a maternal meiosis I non‐disjunction of chromosome 12 in DNA from a cultured placental biopsy. Although predicting the outcomes of pregnancies involving confined placental mosaicism remains difficult, molecular techniques are valuable tools for distinguishing uniparental from biparental disomy and mechanisms of mosaicism.
Prenatal diagnosis of herlitz junctional epidermolysis bullosa by amniocentesis
Authors: Marinkovich M. Peter , Meneguzzi Guerrino , Burgeson Robert E. , Blanchet‐Bardon Claudine , Holbrook Karen A. , Smith Lynne T. , Christiano Angela M. , Ortonne Jean‐Paul
Published: March 2007

Herlitz junctional epidermolysis bullosa (HJEB) is a severe blistering disorder which usually results in death during infancy. We have previously shown that the anchoring filament protein laminin‐5 (kalinin/nicein), which mediates keratinocyte attachment and dermal–epidermal cohesion, is abnormally expressed in individuals with HJEB. Laminin‐5 was detected by Western blot analysis in amniotic fluid from 44 consecutive normal secondtrimester control pregnancies, but was undetectable in second‐trimester amniotic fluid from four pregnancies with fetuses affected by HJEB. In one case of severe non‐Herlitz JEB, laminin‐5 was detected in both amniotic fluid and skin. In human amniotic fluid, the laminin‐5 a3 subunit was processed to a major 165 kD species and a minor 145 kD species and the β2 subunit was partially processed to 105 kD. Although laminin‐5 was covalently associated with laminin‐6 (K‐laminin) in amniotic membrane, no covalent interaction was detected in amniotic fluid. Laminin‐5 from amniotic fluid strongly supported keratinocyte attachment. These results suggest that Western blot analysis of second‐trimester amniotic fluid is useful in determining the prenatal diagnosis of HJEB and that laminin‐5 may serve a physiologically important function in amniotic fluid.
Fetal urinary insulin‐like growth factor I and binding protein 3 in bilateral obstructive uropathies
Authors: Bussieres L. , Laborde K. , Souberbielle J. C. , Muller F. , Dommergues M. , Sachs C.
Published: March 2007

Fetal urinary concentrations of insulin‐like growth factor I (UIGF‐I) and binding protein 3 (UIGFBP‐3) were determined in patients with prenatal diagnosis of bilateral obstructive uropathy. Patients were retrospectively assigned to three groups, on the basis of outcome: group 1, termination of pregnancies (n = 11) with sonographic evidence of severe oligohydramnios or renal dysplasia, confirmed at histological examination; group 2, patients (n = 10) with postnatal plasma creatinine > 50 μmol/1 at the age of 1 year (1 yr‐pCreat); and group 3, patients (n = 16) with 1 yr‐pCreat ≤ μmol/1. The results show a significant increase in UIGF‐I and UIGFBP‐3 in groups 1 (18 159 ± 9083 pg/ml; 2657 ± 669 ng/ml) and 2 (1574 ± 847 pg/ml; 176 ± 50 ng/ml) in comparison with group 3 (35 ± 6 pg/ml; 21 ± 2 ng/ml). UIGF‐I and UIGFBP‐3 were significantly correlated with postnatal plasma creatinine, and were both sensitive (90 per cent; 80 per cent) and specific (88 per cent; 88 per cent) for prediction of elevated 1 yr‐pCreat (>50 μmol/1). Fetal urinary IGF‐I and IGFBP‐3 are increased in severe fetal bilateral obstructive uropathy, possibly reflecting tubular dysfunction or/and increased synthesis consequent upon fetal kidney injury. Their predictive value for postnatal renal function needs further assessment.
Prenatal analysis of the insulin receptor gene in a family with leprechaunism
Authors: Longo Nicola , Langley Sharon D. , Still Maria J. , Elsas Louis J.
Published: March 2007

Leprechaunism is an autosomal recessive disease characterized by intrauterine and postnatal growth restriction, loss of glucose homeostasis, and severe insulin resistance. This disease is caused by a failure of function of the insulin receptor and is lethal early in life. Here we report the prenatal diagnosis of leprechaunism in one consanguineous family, Atl‐1, in which two homozygous‐affected siblings died with leprechaunism. The mutation in their insulin receptor impaired insulin binding and altered receptor signalling. Prenatal diagnosis could not be accomplished using insulin binding to cultured amniocytes, but was possible using mutational analysis of the insulin receptor gene in DNA from amniotic cells.
Trisomy 16 fetus surviving into the second trimester
Authors: Cusick William , Bork Michael , Fabri Betsy , Benn Peter , Rodis John F. , Buttino Louis
Published: March 2007

A 27‐year‐old gravida 4, para 3 was found to have anhydramnios at 14 weeks' gestation following a size/date discrepancy noted at her routine prenatal visit. A detailed ultrasound revealed multiple fetal anomalies including congenital heart defect, chest hypoplasia, and bilateral dysplastic kidneys. Karyotype revealed trisomy 16 in 15/15 cells from a tissue specimen obtained from the fetal cord insertion site following elective pregnancy termination.
Prenatal identification of i(YP) by molecular cytogenetic analysis
Authors: Wang Boris B. T. , Yu Loh‐Chung , Peng Willow , Falk Rena E. , Williams John
Published: March 2007

An i(Yp) is a rare marker chromosome. We present a case of de novo 46,X,i(Yp) detected prenatally in an amniotic fluid specimen. Fluorescence in situ hybridization (FISH) studies using a panel of Y‐specific biotinylated DNA probes identified the marker chromosome as i(Yp). Comparative genomic hybridization (CGH) studies further confirmed the diagnosis. Upon pregnancy termination, external examination of the fetus revealed a generally well‐developed male fetus with slight facial dysmorphism and prominent rocker‐bottom feet. The molecular cytogenetic data in this case proved very useful in genetic counselling and served as a good example illustrating the important role of molecular techniques for accurate identification of marker chromosomes.
Elevated alpha‐fetoprotein and human chorionic gonadotropin as a marker for placental trisomy 16 in the second trimester?
Authors: Zimmermann Roland , Lauper Urs , Streicher Anja , Huch Renate , Hugh Albert
Published: March 2007

In a series of 2961 consecutive cases with second‐trimester biochemical triple screening for Down's syndrome and neural tube defect (NTD), ten (0.3 per cent) showed an apparent increased risk for both conditions. Three cases had chromosomal abnormalities, namely trisomy 16 confined to the placenta. Since placental trisomy 16 as well as cases with increased alpha‐fetoprotein (AFP) and human chorionic gonadotropin (hCG) are associated with (intrauterine growth retardation (IUGR), oligohydramnios, and fetal demise, at least some cases with this atypical biochemical profile could be explained by this chromosomal abnormality. From our results we recommend that in cases with increased risk for both Down's syndrome and NTD, fetal karyotyping should preferably be done on a placental biopsy, especially when ultrasound in the absence of anomalies demonstrates early IUGR.
Repeat maternal serum testing for down's syndrome screening using multiple markers with special reference to free α and free β‐hCG
Authors: Hackshaw A. K. , Densem J. , Wald N. J.
Published: March 2007

To determine the effect of routine repeat testing in serum screening for Down's syndrome, we compared estimates of the detection and false‐positive rates. Five serum markers were measured‐alpha‐fetoprotein (AFP), unconjugated oestriol (uE3), human chorionic gonadotrophin (hCG), and its two subunits, free α and free β‐hCG. First and repeat test marker levels were available from 142 women whose samples had been routinely collected and stored in an antenatal serum bank. Different repeat testing policies were compared for various combinations of the markers. If all women had repeat tests using the four markers AFP, uE3, and free α and free β‐hCG, the detection rate for a 5 per cent false‐positive rate was 69 per cent compared with 65 per cent if no women were repeated. Policies of repeating selected women gave similar results. The small gain in screening performance with repeat testing performed routinely is not worthwhile. If a woman does happen to have a repeat test, her risk estimate should, however, be based on both results, not just the second.
Enhanced twin pregnancy detection within an open neural tube defect and down syndrome screening protocol using free‐beta hCG and AFP
Authors: Barnabei Vanessa M. , Krantz David A. , Macri James N. , Larsen John W.
Published: March 2007

We have applied our multimarker approach of maternal serum alpha‐fetoprotein (AFP) and free‐beta human chorionic gonadotropin (hCG) for Down syndrome screening to multiple gestations to assess its efficacy for improved detection of twin and triplet pregnancies. This study matched 225 cases of twin pregnancy and 39 cases of triplet pregnancy each with ten singleton pregnancies based on gestational week, race, time to receive sample, time of year of sample, and geographical area. The ratios of the MOM for each group at the tenth, 50th, and 90th percentiles were compared by the Wilcoxon test. Risks for twins were calculated using Bayes' rule, the age‐related incidence of twins, and the levels of AFP and free‐beta hCG. The tenth, 50th, and 90th percentiles of free‐beta hCG MOMs in twin and triplet cases were 0.85, 1.99, and 4.51, and 1.38, 2.78, and 4.07, respectively. For AFP, the MOMs at these percentiles were 1.26, 1.91, and 2.99, and 2.02, 2.68, and 5.30, respectively. The twin and triplet distributions for each marker were statistically significantly different from the singleton distributions (P<0.0001) and from each other (P=0.0012). At a twin risk cut‐off of 1 in 50, 77.4 per cent of all twin gestations can be detected in a second‐trimester AFP and free‐beta hCG screening protocol with 5.1 per cent of singleton pregnancies falsely identified as at risk for twins. Our dual marker protocol for mid‐trimester pregnancy screening combining AFP and free‐beta hCG can identify over 77 per cent of twin pregnancies in women less than 35 years of age. This benefit may contribute to an improved outcome of pregnancy by early detection of multiple gestation.
Confined placental mosaicism in term placentae: Analysis of 125 cases
Authors: Artan S. , Başaran N. , Hassa H. , Özalp S. , Şener T. , Şayli B. S. , Cengiz C. , Özdemi̊ M. , Durak T. , Dölen I. , Özgünen T. , Tuna M.
Published: March 2007

In order to determine the incidence of confined placental mosaicism (CPM) in term placentae and to show the presence of specific sites and the effect on fetal development, 125 placentae from uneventful pregnancies were analysed by cytogenetic methods. The incidence was at least 4.8 per cent and there were no specific sites on the placenta. Although the number of cases is still too small, we found CPM to be associated with intrauterine growth retardation in six cases.
First‐trimester diagnosis of Meckel–Gruber syndrome by transabdominal sonography in a low‐risk case
Authors: Braithwaite Jeffrey M. , Economides Demetrios L.
Published: March 2007

Meckel–Gruber syndrome is a rare autosomal recessive disorder often resulting in neonatal death within a few hours of life. The condition is usually diagnosed ultrasonographically in the second trimester and earlier diagnosis has been mainly confined to high recurrence risk pregnancies. We describe the earliest non‐invasive diagnosis of this condition at 12+2 weeks' gestation in a patient with no previous history using transabdominal ultrasound. Pregnancy termination was declined. Subsequent development of anhydramnios in the mid‐trimester resulted in poor fetal visualization. We propose that first‐trimester diagnosis of this condition is not only possible but preferable, as normal liquor volume facilitates visualization of fetal anatomy.
Recurrent congenital toxoplasmosis in a woman with lupus erythematosus
Authors: D'Ercole Claude , Boubli Léon , Franck Jacqueline , Casta Michel , Harle Jean‐Robert , Chagnon Catherine , Cravello Ludovic , Leclaire Michèle , Blanc Bernard
Published: March 2007

We describe the case of a patient with systemic lupus erythematosus, treated by corticosteroids, who presented during two successive pregnancies with serological reactivation of toxoplasmosis associated with fetal lesions. The first infected fetus died in utero with signs of hydrops. The second fetus was treated in utero with a combination of sulfadoxine and pyrimethamine, administered to the mother, and is now well. The increasing number of immunocompromised pregnant patients with immunity to Toxoplasma gondii may lead to a higher risk of reactivation of maternal toxoplasmosis and congenital infection.
Detection of low‐grade mosaicism in fetal cells isolated from maternal blood
Authors: Bischoff Farideh Z. , Lewis Dorothy E. , Simpson Joe Leigh , Nguyen Dianne D. , Scott Jeffrey , Schober Wendy , Murrell Sarah , Elias Sherman
Published: March 2007

Recovering and analysing fetal erythrocytes from maternal blood is being pursued for non‐invasive prenatal genetic diagnosis. We report the observation of 46, XY/47, XXY mosaicism in fetal cells from a woman whose first‐trimester chorionic villus sampling (CVS) initially showed only 46, XY. Only after exhaustive (500 cells) analysis were four XXY cells found in cultured villi.