Prenatal diagnosis of skeletal dysplasia due to FGFR3 gene mutations: a 9-year experience : prenatal diagnosis in FGFR3 gene.

PURPOSE: Prenatal diagnosis with ultrasound findings compatible with skeletal dysplasia due to FGFR3 mutations over a 9 year period in pregnancies and abortuses. METHODS: 54 samples were studied. Aneuploidy studies were carried out on all samples. By sequencing analysis, we determined mutations for achondroplasia (ACH), hypochondroplasia (HCH), and type I and type II tanathophoric dysplasia (TD). RESULTS: 2 chorionic villi samples had a G380R mutation due to a mother with ACH; 4 amniotic fluid samples with TDs in which the foetuses had micromelia plus hypoplastic thoraces; 5 samples from abortuses with TDs. Neither ACH nor HCH occurred in sporadic cases. CONCLUSIONS: Molecular studies in ongoing pregnancies are indicated in cases with an affected parent, a family history with positive molecular studies (maternal anxiety), and when the US finding demonstrates micromelia with a hypoplastic thorax. A protocol for tissues of abortuses should include an X-ray, pathologic anatomy, and genetic studies.

Traslucencia nucal aumentada y cariotipo normal. ¿Qué hacer? / No disponible

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Diagnóstico prenatal no invasivo en células fetales obtenidas de sangre periférica materna / Non-invasive prenatal diagnosis on fetal cells from maternal blood

El aislamiento de los eritroblastos fetales presentes en la sangre materna para su posterior estudio representa un prometedor método de diagnóstico prenatal no invasivo.En nuestro laboratorio hemos realizado un estudio poblacional con el que se pretendía realizar una valoración práctica de las técnicas desarrolladas para un diagnóstico prenatal no invasivo. Para el enriquecimiento de las muestras en eritroblastos fetales se empleó el método que se consideró más adecuado para la rutina del laboratorio y para su posterior estudio mediante técnica de FISH. El estudio permitió determinar que la mayor sensibilidad diagnóstica se obtenía en la semana 15 de gestación (76 por ciento), observándose una clara diferencia entre la sensibilidad obtenida en el primer (25 por ciento) y segundo (61,5 por ciento) trimestre de gestación. Distintos grupos están trabajando para modificar las técnicas actuales y así obtener mejores resultados. Sin embargo, desde el punto de vista de una unidad de diagnóstico prenatal existen todavía aspectos que han de resolverse: - No hay un consenso acerca de la mejor semana para realizar el estudio.- Las técnicas son laboriosas y no recuperan un número suficiente de células para el estudio.- La sensibilidad alcanzada no es óptima. - E I coste es muy caro. Por todo ello esta técnica no puede aplicarse todavía a la rutina del diagnóstico prenatal. En el presente artículo, además de presentar nuestros resultados, se discute sobre el estado actual del tema, así como las perspectivas futuras (AU)

ADN fetal en plasma de gestante: aplicaciones en el diagnóstico prenatal / Fetal DNA in maternal plasma: applications in prenatal diagnosis

La presencia de ADN fetal en el suero y el plasma de gestante está llevando al desarrollo de diferentes estrategias para realizar diagnóstico prenatal no invasivo. Hasta el momento diversos autores han publicado principalmente resultados en la detección de sexo fetal y factor RhD. Estos datos han motivado que nuestro grupo evalúe esta metodología para su posible aplicación diagnóstica. Hemos obtenido resultados satisfactorios en la determinación de sexo fetal, enfermedades mendelianas de origen paterno (como fibrosis quistica y corea de Huntington), así como en la determinación del factor RhD fetal (AU)

Huntington disease-unaffected fetus diagnosed from maternal plasma using QF-PCR.

The discovery of fetal DNA in maternal plasma from early pregnancies has led to new opportunities for clinical application. In the last few years there have been numerous reported applications, mainly fetal gender and RhD genotyping. The prenatal diagnosis of some inherited genetic diseases such as Huntington disease is also very frequently required in the prenatal diagnosis routine. We have successfully diagnosed, with a non-invasive procedure, an unaffected HD fetus at the 13th week of gestation using fetal DNA from maternal plasma and the quantitative fluorescent PCR method, which is one of the most sensitive ways to detect fetal DNA in maternal plasma at such an early time of gestation.

Prenatal detection of a cystic fibrosis mutation in fetal DNA from maternal plasma.

OBJECTIVES: Maternal plasma and serum are being used to detect fetal DNA by PCR in order to determine certain conditions such as fetal gender and RhD without invasive procedures. Because of the presence of maternal DNA in plasma, these approaches are limited to paternally inherited disorders or those de novo present in the fetus. We have assessed the possibility of performing the detection of a single-gene disorder such as a fetal paternally inherited Cystic Fibrosis mutation (Q890X) in maternal plasma. METHODS: The analysis was performed at 13 weeks of gestation using DNA extracted from maternal plasma. We used a PCR amplification of the Q890X mutation and a posterior restriction analysis of the PCR product. RESULTS: We were able to detect the presence of the mutation and thus the fetal condition of being a carrier of the paternal mutation. CONCLUSIONS: We have made evident the possibility of detecting an inherited paternal mutation in a non-invasive way at the 13t(hr) weeks of pregnancy. This methodology could be very useful in cases of paternally inherited dominant disorders. The technical improvements in fetal DNA detection and analysis might lead to the development of new applications in the non-invasive prenatal diagnosis field.

Prenatal diagnosis on fetal cells from maternal blood: practical comparative evaluation of the first and second trimesters.

Objectives- Several attempts have been made to determine the gestational period in which the maximum number of fetal cells can be found in maternal blood and consequently which is the best week in which to perform a reliable non-invasive prenatal diagnosis. Most studies conclude that the number of nucleated red blood cells (NRBC) increases in line with gestation, but the number of cells that are fetal in origin (FNRBC) decreases in the third trimester. The aim of the present study was to make a practical comparative evaluation of the first and second trimesters to ascertain the period in which a greater number of FNRBC can be found of the total number of NRBC identified. Methods- Double density gradient and a posterior positive selection (CD71) by magnetic activated cell sorting (MACS) were employed. In the final fraction, erythroblasts were identified using Kleihauer staining and were studied using the fluorescence in situ hybridization (FISH) interphasic technique. Results- There was a significant difference (p<0.05) between the mean number of FNRBC found in the first and second trimesters. Conclusions- The number of FNRBC increases from the first to the second trimester. It appears that the optimum week in which to perform a reliable non-invasive prenatal diagnosis is around the 15th week.

[Practice Guidelines of the Spanish Society of Cardiology for the management of cardiac disease in pregnancy]. / Guías de práctica clínica de la Sociedad Española de Cardiología en la gestante con cardiopatía.

Maternal adaptation to pregnancy includes reproductive hormone interaction plasma, volume changes with an increase in total body water, vascular alterations with a decrease in systemic resistance and modifications associated with hypercoagulability. These explain, in part, the appearance of signs and symptoms, even in a normal pregnant woman, that are difficult to distinguish from those occurring in heart disease and why some cardiac abnormalities are not well tolerated during pregnancy. Cardiovascular abnormalities are considered the first non-obstetric cause of morbidity and mortality during pregnancy. Rheumatic and congenital heart diseases are currently the most frequent cardiopathy found in women of childbearing age, followed by hypertension, coronary artery disease and arrhythmia. Although pregnancy is well tolerated by most women with heart disease, there are some cardiovascular abnormalities which place the mother and the infant at extremely high risk: patients with congestive heart failure and severe cardiac dysfunction, pulmonary hypertension, cyanotic congenital heart disease, Marfan's syndrome, severe obstructive lesions of the left side of the heart, patients with prosthetic cardiac valves and antecedents of peripartum cardiomyopathy should be encouraged to avoid pregnancy and the interruption of pregnancy may be advisable in cases with great risk of disability or death. The most severe cardiopathies significantly increase the risk of fetal loss and the presence of a congenital cardiac abnormality in either parent increases the risk of congenital cardiac disease in the fetus. Medical care must be initiated early, prior to conception and women with cardiopathy should be informed of the possible risks of pregnancy to both the mother and fetus.

Sonographic, cytogenetic and DNA analysis in four 69,XXX fetuses diagnosed in the second trimester.

OBJECTIVE: To describe the ultrasound findings and its relationship with the cytogenetic study and the origin of the extra haploid chromosome set in four 69,XXX cases. METHODS: Four pregnant women were referred because of abnormal 2nd trimester ultrasound. Karytoypes, FISH and DNA analysis were performed. RESULTS: All cases presented asymmetrical intrauterine growth retardation, marked oligohydramnios and placental alterations and showed a 69,XXX karyotype. In three cases, DNA analysis allowed to establish the origin of the extra haploid chromosome set. CONCLUSIONS: At least three fetuses had a maternal extra haploid chromosome set. Thus, it has been possible to establish the main ultrasonographic markers and to observe the survival of the fetus until the second trimester when they have a maternal origin.

17 Beta-estradiol-stimulated nitric oxide production by neutrophils: effect on platelet activation.

OBJECTIVE: To evaluate the effect of 17beta-estradiol (E2) on the ability of human neutrophils to produce nitric oxide (NO) and its effects on platelet activation. METHODS: The expression of neuronal nitric oxide synthase (nNOS) protein and the formation of NO by 17beta-E2-incubated neutrophils from men were studied in vitro (ten male volunteers, no medical-surgical antecedents, aged 25-45 years). Platelet aggregometry and changes in cyclic guanosine monophospate (cGMP) levels were used to bioassay the functionality of NO released from neutrophils. RESULTS: Incubation of neutrophils derived from men with physiologic concentrations of 17beta-E2 (10(-10) to 10(-8) mol/L) enhanced the expression of nNOS protein. 17Beta-E2-incubated neutrophils also showed a significant increase in their ability to generate NO measured by the conversion of [3H]-L-arginine to [3H]-L-citrulline. Furthermore, 17beta-E2-incubated neutrophils showed a greater ability to prevent adenosine diphosphate (ADP)-induced platelet activation. Moreover, increased levels of cGMP were found in the coincubation of platelets with 17beta-E2-treated neutrophils. CONCLUSION: These results suggest that 17beta-E2 increases the ability of human neutrophils to produce NO and therefore may contribute to cardiovascular disease protection.

Estrogen stimulates neuronal nitric oxide synthase protein expression in human neutrophils.

Recent studies have postulated the contribution of nitric oxide (NO) released by the endothelium to the beneficial effects of estrogen. Despite a neuronal-type NO synthase (nNOS) described in neutrophils, less is known about the effect of estrogen in these cells. The aim of the present study was to analyze the expression of nNOS protein in human neutrophils under different estrogenic conditions. We first analyzed nNOS expression in neutrophils obtained from premenopausal women. During the first 2 days of the follicular phase (low circulating estrogen concentrations), nNOS expression in neutrophils was reduced with respect to that found in neutrophils obtained from the same donors during the ovulatory phase (high circulating estrogen concentrations). Moreover, the expression of nNOS protein in neutrophils obtained from postmenopausal women after transdermal estrogen therapy was markedly enhanced with respect to that observed before the treatment. In vitro incubation of neutrophils derived from men for 6 hours with 17beta-estradiol (10(-10) to 10(-8) mol/L) upregulated the expression of nNOS protein. The 17beta-estradiol receptor antagonists, tamoxifen (10(-8) mol/L) and ICI 182780 (10(-8) mol/L), inhibited the upregulation of nNOS protein induced by 17beta-estradiol. The putative functional implication was denoted by a reduced expression of the CD18 antigen on the surface of 17beta-estradiol-incubated neutrophils, which was accompanied by a decreased adhesive capacity. Both effects were prevented by an NO antagonist. In conclusion, the in vivo levels of circulating estrogen concentrations seem to be associated with the level of nNOS protein expression in neutrophils from women. Moreover, low doses of 17beta-estradiol upregulate nNOS protein expression in neutrophils from men. The increased ability of 17beta-estradiol-incubated neutrophils derived from men to produce NO reduced their adhesive properties.

Prenatal diagnosis on fetal cells obtained from maternal peripheral blood: report of 66 cases.

The potential use of fetal cells circulating in maternal blood for a non-invasive prenatal diagnosis has been widely described. Several authors have developed different methods for the enrichment of fetal cells from maternal peripheral blood. The aim of this study was to make a practical valuation of this new prenatal diagnosis technique, using those methods described as efficient and easy to carry out in a prenatal diagnosis unit. These methods consist of the double-density gradient and the positive selection by magnetic activated cell sorting (MACS) of the fetal erythroblasts, and the posterior study of the cells applying the FISH interphasic technique. Once the technique was ready, we obtained results from the study of 66 venous blood samples from women coming for prenatal diagnosis. Using a specific staining for fetal haemoglobin, fetal cells were identified in 63 cases. Fetal sex was well determined in 56 cases, 23 females and 33 males; in 7 cases the sex determination failed. All the aneuploidies found in a previous prenatal diagnosis were confirmed.

Segregation of digital number with partial monosomy or trisomy of 13q in familial 5;13 translocation.

It has been postulated that the deletion of band 13q22 may be associated with digital malformations, especially thumb and big toe anomalies. We report a family where the mother is carrying a balanced translocation between chromosomes 5p15 and 13q22. The offspring have a specific and well-defined phenotype depending on which is the unbalanced chromosome in the karyotype. When a partial trisomy of 13q22-->qter is present, the fetuses have polydactyly in the four limbs, and when the fetus is carrying a partial monosomy of this portion, an oligodactyly in all members can be observed.

[Evaluation on non invasive diagnostic tests for the second look in epithelial ovarian cancer]. / Valoración de las pruebas diagnósticas no invasivas frente al second look en el cáncer epitelial de ovario.

OBJECTIVE: To determine the usefulness of diagnostic tests performed before a second look laparotomy in patients with epithelial ovarian cancer. STUDY DESIGN: Thirty-three patients with epithelial ovarian cancer attended at Fundación Jiménez Díaz from 1984 to 1995 were studied. All patients initially underwent cyto-reducing surgery, followed by at least six platinum-based chemotherapy cycles. Prior to second look laparotomy all patients were evaluated by computerized tomography (CT) of the pelvis and abdomen, CA-125, pelvic-abdominal echography and gynecologic examination. To evaluate sensitivity, specificity, positive predictive value and negative predictive value for each test contingency tables were used. RESULTS: Eleven out of the 33 second look patients (33%) had histologic or cytologic evidence of disease. Six out of the eleven positive second look had a positive CT prior to second look (sensitivity of 55%). CT showed lack of disease in 21 out of the 22 negative second look cases (specificity 95%). Positive and negative predictive values of the test were 86% and 81%, respectively. Nine cases out of the 28 who had a CA-125 obtained had a positive second look. Four out of these nine patients had an increased CA-125 value (sensitivity 44%, specificity 95%, positive predictive value 80% and negative predictive value 78%). Sensitivity, specificity, positive predictive value and negative predictive value of physical examination and echography were 36%, 100%, 100%, 76% and 27%, 95%, 75%, 72%, respectively. On the other hand, sensitivity, specificity, positive predictive value and negative predictive value of all tests taken together were 64%, 91%, 78% and 83%, with a rate of false-negative results of 17% and a rate of false-positive results of 22%. CONCLUSION: Pelvic-abdominal computerized tomography, CA-125, pelvic-abdominal echography and gynecologic examination can be an alternative to second look laparotomy for the diagnosis of persistence or recurrence of the disease in patients with epithelial ovarian cancer.

Ultrasonographic appearance of an echinococcus ovarian cyst.

BACKGROUND: We describe the ultrasonographic pattern of a pelvic echinococcus cyst, visualized using a vaginal probe and color Doppler. CASE: A 27-year-old woman presented with an asymptomatic right adnexal mass. Vaginal ultrasonography revealed a cyst in the posterior cul-de-sac, adjacent to the right ovary, with internal septae resembling a maze or an onion slice structure. Color Doppler revealed peripheral vascularization with a low resistance pattern (resistance index 0.6; pulsatility index 0.93). Benign cysts usually have a simple echogenic pattern. In contrast, ovarian carcinomas have a complex internal structure. CONCLUSION: Ovarian echinococcus cysts may have a peculiar ultrasonographic pattern at high-frequency (7.5-MHz) vaginal ultrasonography. Such high-frequency ultrasonography can be a useful tool in the diagnosis of the disease.

Antenatal diagnosis and management of a ranula.

We describe a case of congenital ranula or retention salivary cyst diagnosed at 26 weeks of gestation. The fetal karyotype was obtained by amniocentesis and chorionic villus sampling, and was normal. Follow-up ultrasound scans revealed no changes in the size or the position of the cyst. Surgical treatment was performed 2 days after a normal vaginal delivery, with excellent results.

[Intrahepatic cholestasis of pregnancy. A benign disease?]. / Colestasis intrahepática del embarazo. Una enfermedad benigna?

Intrahepatic cholestasis of pregnancy is a relatively common disease. It has an unknown etiology and may have a recurrent pattern. It commonly occurs in the 2nd-3rd trimester and characteristically presents with pruritus, jaundice and abnormal liver function tests. There is also an increased risk of preterm delivery and of cesarean section. Both maternal and neonatal prognosis is generally good. We describe a case of intrahepatic cholestasis of pregnancy with an atypical presentation and outcome. Our patient presented with acute renal and hepatic failure with hepatic encephalopathy, DIC and hypertension which was the cause of the fetal death in the third trimester of the pregnancy.