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La identificación de anomalías cromosómicas fetales es una de las principales tareas a las que debe enfrentarse cualquier obstetra involucrado en el diagnóstico de anomalías congénitas. Se analizaron características clínicas y citogenéticas en gestantes sometidas a amniocentesis. Estudio observacional, descriptivo y retrospectivo, incluyó casos consecutivos en un centro privado de julio de 2010 a enero de 2015. Las muestras fueron procesadas en CGC Genetic (Porto-Portugal). Para el análisis estadístico se utilizó PEPI 4.0X. Se realizaron 67 estudios, en el 98,5% se pudo obtener el cariotipo y de éstos resultaron 74,2% normales y 25,8% anormales: 35,4% Trisomía 21, 17,6% Trisomía 18, Trisomía 13 y Sx Turner respectivamente, entre las principales. Indicaciones: 6,0% edad materna, 14,9% edad materna + alteración ecográfica, 77,6% alteración ecográfica (45,2 malformaciones congénitas mayores, 20,1% translucencia nucal aumentada, 17,7% higroma quístico entre otras). La edad gestacional promedio de la punción fue 19 semanas, la menor a las 15 y la mayor a las 30. El resultado del cariotipo se recibió 12 días posteriores a la técnica en promedio, mínimo 8 días y máximo 29. No se presentaron complicaciones obstétricas. El seguimiento de los casos encontró concordancia entre el cariotipo y el fenotipo del recién nacido. Si bien es un número bajo de muestras, la amniocentesis es un método diagnóstico confiable y de bajo riesgo. El diagnóstico prenatal de cromosomopatías permitió el asesoramiento genético y el manejo obstétrico y pediátrico de los casos de manera adecuada. En los embarazos con cariotipo normal, este resultado alivió la preocupación de muchos padres. Identification of fetal chromosomal abnormalities is one of the main issues which need to be addressed by any obstetrician involved in diagnosis of congenital anomalies. Clinical and cytogenetic characteristics were analyzed in pregnant women subjected to amniocentesis. This was an observational descriptive retrospective study that included consecutive cases in a private center from July, 2010 to January, 2015. The samples were processed in CGC Genetic (Porto - Portugal). For the statistical analysis, PEPI 4.0X. was used. Sixty seven studies were conducted, in 98.5% karyotype was obtained , 74.2% was normal and 25.8% abnormal: 35.4% Trisomy 21, and 17.6% Trisomy 18, Trisomy 13 and Sx Turner respectively. Among the the main indications: 6.0% maternal age, 14.9% maternal age plus ultrasound alteration, 77.6% ultrasound alteration (45.2% major congenital malformations, 20.1 increased nuchal translucency , 17.7% cystic hygroma among others). The mean gestational age of the puncture was 19 weeks, the lowest 15 and the highest 30. The result of the karyotype was received 12 days after the technique, the minimum 8 days and the maximum 29. There were no obstetric complications. The follow-up of cases found concordance between the karyotype and the phenotype of the newborn. Although this was a low number of samples, the amniocentesis was reliable diagnostic method with low risk. The prenatal diagnosis of chromosomopathies allowed the genetic counseling and appropiate obstetric and pediatric management cases. In pregnancies with normal karyotype, this result alleviated the concern of many parents.

Mémoire de DES (Diplôme d'Etudes Spécialisées) tenant lieu de thèse d'exercice.; Male infertility affects more than 20 million men worldwide and represents a major public health issue. Although it involves many factors, male infertility has a significant genetic component. My work consisted in characterizing a novel gene in the phenotype of Multiple Morpholgical Abnormalities of the Flagellum (MMAF), a teratozoospermia responsible for severe malformations of the sperm flagellum. The exome sequencing of a cohort of 73 infertile patients with MMAF has allowed the identification of homozygous truncating mutations in the gene CFAP43 in 5 patients. The study of the transcripts by RT-qPCR showed predominant expression in testis. The sperm analysis by electron microscopy and immunofluorescence as well as knockout mouse production by the technique of CRISPR / case.9 was used to demonstrate the involvement of CFAP43 in the MMAF phenotype of our patients. In addition, one of the therapeutical options for these patients MMAF is to resort to medically assisted reproduction technology (ART) including in intracytoplasmic sperm injection (ICSI). Therefore, the second aim of my thesis was to verify whether mutations in the genes responsible for this phenotype did not influence the chances of success of ICSI in these patients. We retrospectively analyzed the ICSI success rate in 6 patients MMAF mutated gene in DNAH1 and assessed the quality of chromatin gametes. In conclusion, we observed a good pregnancy rate (50%), good quality and low nuclear aneuploidy justifying the medical management of these couples by ICSI.; L’infertilité masculine concerne plus de 20 millions d’hommes à travers le monde et représente un véritable enjeu de santé publique. Bien que multifactorielle, l’infertilité masculine a une composante génétique importante. Mon travail a consisté à caractériser un nouveau gène dans le phénotype de Malformation Multiple des Flagelles (MMAF), une tératozoospermie responsable de malformations sévères du flagelle du spermatozoïde. Le séquençage exomique d’une cohorte de 73 patients infertiles avec MMAF a permis d’identifier des mutations tronquantes homozygotes dans le gène CFAP43 chez 5 patients. L’étude des transcrits par RT-qPCR a démontré une expression majoritaire dans le testicule. Les analyses par microscopie électronique et immunofluorescence des spermatozoïdes de même que la production de souris KO par la technique de CRISPR/Cas9 ont permis de confirmer formellement l’implication du gène CFAP43 dans le phénotype MMAF des patients. Par ailleurs, une des options thérapeutiques chez ces patients MMAF est de recourir à des techniques de procréation médicalement assistée (PMA) et notamment à l'injection intracytoplasmique de spermatozoïdes (ICSI). Le second objectif de ma thèse a été de vérifier si les mutations dans les gènes responsables de ce phénotype n’entravaient pas les chances de réussite de l’ICSI chez ces patients. Nous avons analysé rétrospectivement le taux de réussite de l'ICSI chez 6 patients MMAF mutés dans le gène DNAH1 et évalué la qualité de la chromatine de leurs gamètes. Au final, on note un bon taux de grossesses (50%), une bonne qualité nucléaire et un faible taux d'aneuploïdie justifiant la prise en charge médicale par ICSI de ces couples.

Antecedentes: El cribado combinado de cromosomopatías, fundamentalmente trisomía 21, en el primer trimestre de gestación, se introdujo en los años '90 y está basado en un cálculo del riesgo a través de la combinación de la edad materna, la translucencia nucal fetal (TN), y los marcadores bioquímicos maternos (PAPP-A y fBHCG). Mediante esta combinación de marcadores se obtiene una sensibilidad para aneuploidías del 85-90%, con una tasa de falsos positivos (FP) del 5%. Objetivo: El propósito de este trabajo es describir la población cribada y analizar los resultados desde la implantación de la técnica en nuestro centro. Método: Estudio observacional de la población gestante que acudió para cribado de cromosomopatías durante el primer trimestre de la gestación al Hospital Clínico San Carlos, desde julio de 1999 hasta diciembre de 2009. Resultados: El tamaño muestral fue de 21.194 gestaciones simples, con un 12,5% de mujeres con más de una gestación y 141 casos de aneuploidía (6,6%). Se describe las características de la población y el comportamiento de las variables estudiadas. La sensibilidad del cribado combinado fue del 70%, con una tasa de FP de 2,3%. Conclusión: La tasa de detección del cribado combinado de cromosomopatías en el primer trimestre de gestación, es menor que la publicada en la literatura, aunque ha ido mejorando con los años, probablemente por una mejor formación de los ecografistas. En contraposición, la tasa de FP es muy baja. Background: Screening for aneuploidies, mainly trisomy 21, during the first trimester of gestation, was introduced in the '90s and is based on a calculation of the risk through the combination of maternal age, nuchal translucency (NT) and biochemical parameters (PAPP-A and fBHCG). By means of this combination we can obtain a detection rate for aneuploidies of 85-90%, with a false positive rate of 5%. Objective: To describe our population and analyze our results related to combined screening for aneuploidies, during the period of time it has been performed in our hospital. Methods: Observational study of all pregnant women who attended Hospital Clínico San Carlos for screening of chromosomopaties during first trimester of gestation, from July 1999 to December 2009. Results: Sample size was 21,194 single pregnancies, with 12,5% of women with more than one gestation, and 141 cases of aneuploidy (6,6%). We describe the characteristics of our population and the distribution of the parameters studied. Combined screening had a detection rate of 70% with a false positive rate of 2,3%. Conclusion: Screening for aneuploidies during first trimester of gestation, in Hospital Clínico San Carlos, has a lower detection rate than previously reported. However, the false positive rate is very low.

Sister chromatid cohesion is essential for cell division. During meiosis, it is also required for proper synapsis of pairs of sister chromatids and for chiasma formation and maintenance. Since mammalian oocytes remain arrested in late prophase for a very long period—up to five decades in humans—the preservation of cohesion throughout this period is a formidable challenge. Mouse models with cohesin deficiencies and aging wild-type mice showed that this challenge is not fully met: cohesion weakens and deteriorates with increasing age. These recent findings have highly significant implications for our comprehension of the genesis of aneuploidies.

Background: Little is known about the physical and mental health status among parents of boys with SCA. The term sex chromosome aneuploidies (SCA) refers to a condition where children are born with additional sex chromosomes. SCA in boys may be associated with physical, psychological, cognitive, and interpersonal challenges. Boys with SCA are at increased risk of motor- and language developmental delays, learning disabilities, social difficulties and behavioral problems. Objectives and methods: To investigate subjective health complaints and personal well-being (PWB) in a sample of parents (n = 38) to boys with SCA recruited from a database of families from Frambu Resource Center for Rare Disorders and the annual meeting of the National Klinefelter Association in Norway. Parents received envelops with demographic questions and questionnaires that included the Subjective Health Complaints Inventory (SHC) and the Personal Wellbeing Index - Adult (PWI-A). Results: The prevalence of subjective health complaints in the parent sample was high. The parents reported low health satisfaction on the PWI-A, but total PWI-A was high. Mothers in the parent sample reported higher frequencies on subjective health complaints on the SHC compared to fathers, as well as lower health satisfaction on the PWI-A. A negative association between health satisfaction on the PWI-A and the SHC was found, as well as between pseudoneurological complaints on the SHC and the total PWI-A. Conclusion: Despite the high amount of subjective health complaints and low health satisfaction found among parents of boys with SCA, PWB was generally high. The findings support previous research on parents of children with disabilities and chronic disorders. Future studies should compare parents of boys with SCA to parents of children with other disabilities, and investigate differences in functioning among parents of boys with different SCA karyotypes. Information about gender differences and coping among parents of children with SCA might be helpful to relieve burden, and more effort should be put in to help identify parents in need of interventions.

To investigate large structural clonal mosaicism of chromosome X, we analysed the SNP microarray intensity data of 38,303 women from cancer genome-wide association studies (20,878 cases and 17,425 controls) and detected 124 mosaic X events >2 Mb in 97 (0.25%) women. Here we show rates for X-chromosome mosaicism are four times higher than mean autosomal rates; X mosaic events more often include the entire chromosome and participants with X events more likely harbour autosomal mosaic events. X mosaicism frequency increases with age (0.11% in 50-year olds; 0.45% in 75-year olds), as reported for Y and autosomes. Methylation array analyses of 33 women with X mosaicism indicate events preferentially involve the inactive X chromosome. Our results provide further evidence that the sex chromosomes undergo mosaic events more frequently than autosomes, which could have implications for understanding the underlying mechanisms of mosaic events and their possible contribution to risk for chronic diseases., It is unclear how often genetic mosaicism of chromosome X arises. Here, the authors examine women with cancer and cancer-free controls and show that X chromosome mosaicism occurs more frequently than on autosomes, especially on the inactive X chromosome, but is not linked to non-haematologic cancer risk

STUDY QUESTION Is blastocyst biopsy and quantitative real-time PCR based comprehensive chromosome screening a consistent and reproducible approach across different biopsy practitioners? SUMMARY ANSWER The blastocyst biopsy approach provides highly consistent and reproducible laboratory and clinical outcomes across multiple practitioners from different IVF centres when all of the embryologists received identical training and use similar equipment. WHAT IS KNOWN ALREADY Recently there has been a trend towards trophectoderm (TE) biopsy in preimplantation genetic screening (PGS)/preimplantation genetic diagnosis (PGD) programmes. However, there is still a lack of knowledge about the reproducibility that can be obtained from multiple biopsy practitioners in different IVF centres in relation also to blastocysts of different morphology. Although it has been demonstrated that biopsy at the blastocyst stage has no impact on embryo viability, it remains a possibility that less experienced individual biopsy practitioners or laboratories performing TE biopsy may affect certain outcomes. We investigated whether TE biopsy practitioners can have an impact on the quality of the genetic test and the subsequent clinical outcomes. STUDY DESIGN, SIZE, DURATION This longitudinal cohort study, between April 2013 and December 2014, involved 2586 consecutive blastocyst biopsies performed at three different IVF centres and the analysis of 494 single frozen euploid embryo transfer cycles (FEET). PARTICIPANTS/MATERIALS, SETTING, METHODS Seven biopsy practitioners performed the blastocyst biopsies in the study period and quantitative PCR was used for comprehensive chromosome screening (CCS). The same practitioner performed both the biopsy and tubing procedures for each blastocyst they biopsied. To investigate the quality of the biopsied samples, the diagnostic rate, sample-specific concurrence and the cell number retrieved in the biopsy were evaluated for each biopsy operator. Clinical outcomes following FEET cycles were stratified by biopsy operator and compared. Cellularity of the biopsy sample was also correlated with clinical outcomes. MAIN RESULTS AND THE ROLE OF CHANCE The seven practitioners performed 2586 biopsies, five in centre IVF-1 and one in each of the other two IVF centres (IVF-2 and IVF-3). Overall, 2437 out of 2586 (94.2%) blastocyst biopsies resulted in a conclusive diagnosis, 119 (4.6%) showed a nonconcurrent result and 30 (1.2%) failed to amplify, suggesting the absence of TE cells in the test tube or presence of degenerated/lysed cells only. Among the samples producing a conclusive diagnosis, a mean concurrence value of 0.253 (95% CI = 0.250–0.257) was observed. Logistic regression analysis adjusted for confounding factors showed no differences in the diagnosis rate and in the concurrence of the genetic analysis between different biopsy practitioners. An overall mean number of 7.32 cells (95% CI = 6.82–7.81; range 2–15) were predicted from all biopsies. Higher cellularity was significantly associated with a better quality of the CCS diagnosis (P < 0.01) and with the conclusive diagnosis rate, with nonconcurrent samples showing significantly lower numbers of cells (2.1; 95% CI=1.5–2.7) compared with samples resulting in a conclusive diagnosis (mean cells number 7.5; 95% CI = 7.1–7.9, P < 0.01). However, no differences were recorded between different biopsy practitioners regarding cellularity of the biopsy. Finally, logistic analysis showed no impact of the biopsy practitioners on the observed ongoing rates of implantation, biochemical pregnancy loss and miscarriage after the FEET cycles. LIMITATIONS, REASONS FOR CAUTION These data come from a restricted set of laboratories where all of the embryologists received identical training and use identical equipment. A single TE biopsy method and CCS technology was used and these data particularly apply to PGS programmes using blastocyst biopsy without zona opening at the cleavage stage and using qPCR-based CCS. To make firm conclusions on the potential impact of biopsy on biochemical pregnancy loss and miscarriages according to practitioner and biopsy cellularity, a larger sample size is needed. WIDER IMPLICATIONS OF THE FINDINGS We reported a very high consistency and reproducibility of the blastocyst biopsy approach coupled with qPCR-based CSS for both genetic and clinical outcomes across different practitioners working in different IVF centres when appropriate training is provided and when the same laboratory setting is used. These data are important considering the trend towards the use of blastocyst biopsy worldwide for PGD/PGS applications. STUDY FUNDING/COMPETING INTEREST(S) None.

The objective of this study is to examine the potential value of using machine learning techniques such as artificial neural network (ANN) schemes for the noninvasive estimation, at 11-13 weeks of gestation, the risk for euploidy, trisomy 21 (T21), and other chromosomal aneuploidies (O.C.A.), from suitable sonographic, biochemical markers, and other relevant data. A database(1) consisted of 51,208 singleton pregnancy cases, while undergoing first trimester screening for aneuploidies has been used for the building, training, and verification of the proposed method. From all the data collected for each case from the mother and the fetus, the following 9 are considered by the collaborating obstetricians as the most relevant to the problem in question: maternal age, previous pregnancy with T21, fetal crown-rump length, serum free beta-hCG in multiples of the median (MoM), pregnancy-associated plasma protein-A in MoM, nuchal translucency thickness, nasal bone, tricuspid flow, and ductus venosus flow. The dataset was randomly divided into a training set that was used to guide the development of various ANN schemes, support vector machines, and k-nearest neighbor models. An evaluation set used to determine the performance of the developed systems. The evaluation set, totally unknown to the proposed system, contained 16,898 cases of euploidy fetuses, 129 cases of T21, and 76 cases of O.C.A. The best results were obtained by the ANN system, which identified correctly all T21 cases, i.e., 0% false negative rate (FNR) and 96.1% of euploidies, i.e., 3.9% false positive rate (FPR), meaning that no child would have been born with T21 if only that 3.9% of all pregnancies had been sent for invasive testing. The aim of this work is to produce a practical tool for the obstetrician which will ideally provide 0% FNR and to recommend the minimum possible number of cases for further testing such as invasive. In conclusion, it was demonstrated that ANN schemes can provide an effective early screening for fetal aneuploidies at a low FPR with results that compare favorably to those of existing systems.

Objectives To report clinical implementation of cell-free DNA (cfDNA) analysis of maternal blood in screening for trisomies 21, 18 and 13 in twin pregnancies and examine variables that could influence the failure rate of the test. Methods cfDNA testing was performed in 515 twin pregnancies at 10–28 weeks' gestation. The failure rate of the test to provide results was compared with that in 1847 singleton pregnancies, and logistic regression analysis was used to determine which factors among maternal and pregnancy characteristics were significant predictors of test failure. Results Failure rate of the cfDNA test at first sampling was 1.7% in singletons and 5.6% in twins. Of those with a test result, the median fetal fraction in twins was 8.7% (range, 4.1–30.0%), which was lower than that in singletons (11.7% (range, 4.0–38.9%)). Multivariable regression analysis demonstrated that twin pregnancy, higher maternal weight and conception by in-vitro fertilization provided significant independent prediction of test failure. Follow-up was available in 351 (68.2%) of the twin pregnancies and comprised 334 with euploid fetuses, 12 discordant for trisomy 21 and five discordant for trisomy 18. In all 323 euploid cases with a result, the risk score for each trisomy was

Introducción: El diagnóstico genético preimplantacional (PGD) por medio de las técnicas de aCGH y polimorfismo de nucleótido único (SNPs - single nucleotide polymorphism) se ha convertido en una herramienta útil en los ciclos de reproducción asistida, superando al PGD por hibridación fluorescente in situ (FISH). Estas técnicas nos permiten conocer las aneuploidías en cada uno de los cromosomas. Además, la SNPs con Parental Support (Natera, Inc) posibilita conocer el origen de la aneuploidía en los blastocistos, si por el espermatozoide o por el ovo-cito. Objetivos: Determinar la tasa de aneuplodías únicas en embriones humanos obtenidos por reproducción asistida, utilizando la técnica de polimorfismo de nucleótido único. Diseño: Estudio retrospectivo. Institución: Grupo PRANOR, Sede Monterrico, y Genomics Perú, Lima, Perú. Material biológico: Embriones humanos. Intervenciones: Análisis de los registros de 429 embriones estudiados con PGD por SNPs, embriones obtenidos de 105 ciclos de reproducción asistida, entre 2011 y 2013. Principales medidas de resultados: Aneuploidía de embriones, relación con edad materna y origen paterno o materno. Resultados: El 48,8% de embriones resultó normal, con tasa de aneuploidía de 51,2%. La proporción de embriones sanos varió de acuerdo a la edad de la madre, disminuyendo cuando la edad aumentaba. En todos los grupos etarios estudiados más de 66% de las aneuploidías fue de origen materno, incluyendo el grupo de ovodonación (OD). Conclusiones: El diagnóstico preimplantacional mediante SNPs tendría gran valor pronóstico y sería una herramienta útil para conocer el origen de las aneuploidías en los embriones de las pacientes que se someten a procedimientos de reproducción asistida. Background: Preimplantation genetic screening (PGS) using comparative genomics hybridization (aCGH) and single nucleotide polymorphism (SNP) technology has become a useful tool in assisted reproduction by improving clinical outcomes. Both techniques identify aneuploidies and chromosomal rearrangement. However, SNPs with Parental Support (Natera Inc.) give extra information of the aneuploidy origin either from the sperm or the oocyte. Objectives: To determine single aneuplodies rate in human embryos obtained by assisted reproduction using single nucleotide polymorphism microarray. Design: Retrospective study. Setting: Grupo PRANOR, Sede Monterrico, and Genomics Peru, Lima, Peru. Biologic material: Human embryos. Interventions: From 2011 to 2013, 105 patients underwent IVF/ ICSI with SNPs and Parental Support. In total, 429 embryos were analyzed and records were reviewed. Main outcome measures: Embryos aneuploidy, relation with maternal age and paternal or maternal origin. Results: From the 429 embryos 208 (48.8%) were chromosomally normal. The proportion of normal embryos decreased with increasing maternal age. In all age groups more than 66% of aneuploidies had maternal origin, including the ovodonation group (OD). Conclusions: PGS by SNPs with Parental Support resulted in good prognostic value and would be useful in determining the origin of aneuploidy in embryos of patients who underwent assisted reproduction technology.

Background Supernumerary sex chromosome aneuploidies (sSCA) are characterized by the presence of one or more additional sex chromosomes in an individual’s karyotype; they affect around 1 in 400 individuals. Although there is high variability, each sSCA subtype has a characteristic set of cognitive and physical phenotypes. Here, we investigated the differences in the morphometry of the human corpus callosum (CC) between sex-matched controls 46,XY (N =99), 46,XX (N =93), and six unique sSCA karyotypes: 47,XYY (N =29), 47,XXY (N =58), 48,XXYY (N =20), 47,XXX (N =30), 48,XXXY (N =5), and 49,XXXXY (N =6). Methods We investigated CC morphometry using local and global area, local curvature of the CC boundary, and between-landmark distance analysis (BLDA). We hypothesized that CC morphometry would vary differentially along a proposed spectrum of Y:X chromosome ratio with supernumerary Y karyotypes having the largest CC areas and supernumerary X karyotypes having significantly smaller CC areas. To investigate this, we defined an sSCA spectrum based on a descending Y:X karyotype ratio: 47,XYY, 46,XY, 48,XXYY, 47,XXY, 48,XXXY, 49,XXXXY, 46,XX, 47,XXX. We similarly explored the effects of both X and Y chromosome numbers within sex. Results of shape-based metrics were analyzed using permutation tests consisting of 5,000 iterations. Results Several subregional areas, local curvature, and BLDs differed between groups. Moderate associations were found between area and curvature in relation to the spectrum and X and Y chromosome counts. BLD was strongly associated with X chromosome count in both male and female groups. Conclusions Our results suggest that X- and Y-linked genes have differential effects on CC morphometry. To our knowledge, this is the first study to compare CC morphometry across these extremely rare groups.

Introduction. Aneuploidies are the major cause of perinatal death and early psychophysical disorders. Objective. In this study, we analyzed detection and false-positive rates of screening for aneuploidies in the first trimester by the combination of maternal age, fetal nuchal translucency (NT) thickness and maternal serum free beta-human chorionic gonadotrophin (β-hCG), and pregnancy-associated plasma protein-A (PAPP-A) at 11-13+6 weeks of gestation, using the appropriate software developed by the Fetal Medicine Foundation. Methods. Our screening study for aneuploidies analyzed 4172 singleton pregnancies from January 2006 to December 2010. The sensitivities and false-positive rates using the combined aneuploidies determination for the risk cut-off of 1:275 were evaluated. Results. In the trisomy 21 pregnancies, the fetal NT was higher than 95th centile, in 72.8%, serum free b-hCG concentration it was above the 95th centile in 55% and serum PAPP-A was below the 5th centile in 47% of the cases. In the trisomy 18 and 13, the fetal NT was above 95th centile in 66.6% and 44.4% of the cases, respectively. The serum free b-hCG concentration was above the 95th centile in 0 and 10%, but serum PAPP-A was below 5th centile in 80.9% and 88.8% of pregnancies. In the trisomy 21 pregnancies the median free beta-hCG was 2.3 MoM and the median PAPP-A was 0.45 MoM. Chromosomal abnormalities were detected in 169 fetuses: trisomy 21 (97), Turner syndrome (19), trisomy 18 (28), trisomy 13 (11) and others (14). Detection rate of combined screening for aneuploides were 86.0% with false positive rate of 5.3% (mean age 33±4.9 years, >35 years in 35% of pregnancies). Conclusion. Our study suggests that the strategy of first-trimester combined screening of biochemical values and ultrasonographic parameters at 12 gestational weeks identifies higher percentage of aneuploidies with a lower false-positive rate than a single parameter strategy.

Prenatal diagnosis mainly focuses on revealing aneuploidies - the most frequent cause of mental and physical retardation in the childhood and perinatal death. Nowadays, fetal cells are obtained by invasive methods of prenatal diagnosis with some risk of spontaneous miscarriage. To eliminate those perils only pregnancies detected by screening as presenting high risk of aneuploidies are subjected to the cytogenetic and molecular-genetic methods. Powered by TCPDF (www.tcpdf.org)

Titulo en ingles: Effect of protein level on growth of yaque Leiarius marmoratus (Gill, 1870) under farming conditionsTitulo en portugues: Efeito do nível de proteína no crescimento do yaque Leiarius marmoratus (Gill, 1870), sob condições de cultivoResumen: El yaque Leiarius marmoratus ha sido considerada una especie de fácil adaptación al consumo de alimentos secos y por lo tanto con potencial para acuicultura; sin embargo, su comportamiento productivo bajo condiciones de cultivo aún no ha sido suficientemente evaluado En consecuencia, el objetivo de este trabajo fue estudiar la ganancia total de peso (GP), la ganancia diaria de peso (GDP), la conversión alimenticia (CA), la tasa de crecimiento específica (TCE), el factor de crecimiento relativo (FCR) y la sobrevivencia bajo condiciones de cultivo en estanques de tierra, alimentándolo con tres diferentes niveles de proteína bruta (PB) en la dieta (24%, 30% y 34 %) Para este propósito fueron seleccionados juveniles de 3 5 ± 2 9 g de peso y 6 6 ± 1 5 cm de longitud total y sembrados en estanques en tierra (400 m2aproximadamente) a una densidad de 1 individuo/m2 Se midió mensualmente, durante 150 d de cultivo, el peso corporal (g) y la longitud total (cm) El alimento fue suministrado tres veces al día, a las 07:00, 17:00 y 20:00 horas Al finalizar el ensayo, de tres individuos de cada tratamiento se tomó una muestra de músculo dorsal (c.a. 400 g), la cual fue conservada a -20 ºC para posteriormente determinar los porcentajes de proteína cruda, materia seca, cenizas y extracto etéreo La longitud fue mayor (P

ANTECEDENTS: Alguns pacients infèrtils amb cariotip somàtic normal presenten alteracions en el procés de l’espermatogènesi i increments d’anomalies cromosòmiques numèriques en les cèl·lules germinals. S'ha proposat que les aneuploïdies activen els sistemes de control cel·lular i desencadenen en processos de mort cel·lular programada. OBJECTIUS: 1) Desenvolupar una metodologia seqüencial que permeti la detecció, l’aïllament i l’anàlisi citogenètica de cèl·lules testiculars apoptòtiques. 2) Avaluar la relació entre la presència d’anomalies cromosòmiques i la inducció de processos apoptòtics en cèl·lules germinals prereduccionals, postreduccionals, fases meiòtiques, espermatozoides i cèl·lules de Sertoli procedents de biòpsies testiculars d'individus infèrtils. 3) Analitzar la freqüència d’anomalies cromosòmiques observades en les diferents etapes de l’espermatogènesi i contrastar aquesta informació amb les característiques seminals, l’estudi meiòtic i l’anàlisi de FISH en espermatozoides a fi d’identificar pacients amb un comportament similar. MATERIAL I MÈTODES: Per assolir aquests objectius es van utilitzar 31 biòpsies testiculars de pacients que van consultar per problemes de fertilitat. A partir del protocol desenvolupat es va dur a terme l'anàlisi de les anomalies cromosòmiques numèriques i dels processos apoptòtics. Es va analitzar la constitució cromosòmica de les cèl·lules germinals (pre-reduccionals, figures meiòtiques, cèl·lules post-reduccionals i espermatozoides) i de cèl·lules de Sertoli presents en les poblacions cel·lulars viable, apoptòtica i no seleccionada. Es va comparar la freqüència poblacional d’anomalies cromosòmiques entre les diferents fraccions cel·lulars. Finalment, es va realitzar un anàlisi d'agrupació basant-nos en els resultats obtinguts per cada individu. RESULTATS: Els resultats obtinguts demostren que la metodologia d’anàlisi és eficient per detectar l’associació de processos apoptòtics i anomalies cromosòmiques en cèl·lules testiculars. L’anàlisi dels resultats ha posat de manifest una selecció de les anomalies cromosòmiques cap a les fraccions apoptòtiques en les cèl·lules germinals pre-reduccionals i en els espermatozoides. En les cèl·lules germinals post-reduccionals, cèl·lules en profase I i cèl·lules de Sertoli ambdós processos són independents. CONCLUSIONS: 1) La metodologia desenvolupada permet la detecció, l'aïllament i l’anàlisi citogenètica de cèl·lules apoptòtiques en les diferents fases de l’espermatogènesi i en les cèl·lules de Sertoli de biòpsies testiculars diagnòstiques. 2) Les aneuploïdies observades en cèl·lules prereduccionals s’originen per errors de segregació durant la proliferació mitòtica i són eficientment eliminades via apoptosi. 3) La població d’espermatòcits primaris en profase I és majoritàriament euploide i no s'observa una relació entre la presència d'aneuploïdies i els processos apoptòtics. 4) No s'ha trobat correlació entre la presència d’aneuploïdies i d'apoptosi a les cèl·lules de Sertoli. 5) El punt de control del SAC no és eficient, fet que es reflecteix en un increment d’aneuploïdies en cèl·lules postreduccionals. No s'ha observat correlació entre la presència d'aneuploïdies i processos apoptòtics en cèl·lules postreduccionals. Aquestes cèl·lules són eliminades per vies independents de processos apoptòtics. 6) S'ha observat una relació entre els processos apoptòtics i la presència d'anomalies cromosòmiques en els espermatozoides. 7) La població analitzada és molt heterogènia pel que fa als percentatges d'anomalies cromosòmiques i a les dades clíniques. La qual cosa no ha permès determinar característiques clíniques comunes en les agrupacions realitzades en funció dels percentatges d’anomalies cromosòmiques en els diferents etapes de l’espermatogènesi. Aquestes dades reflecteixen la diversitat citogenètica i fenotípica de les poblacions infèrtils., BACKGROUND: Some infertile patient with normal somatic karyotype displays spermatogenic disorders and increased rates of numerical chromosome abnormalities in germ cells. It has been proposed that aneuploidies triggers cell cycle checkpoints leading to apoptosis. OBJECTIVES: 1) To develop a methodology allowing the sequential detection, isolation and cytogenetic analysis of apoptotic testicular cells. 2) To assess the relationship between the presence of chromosomal abnormalities and induction of apoptotic processes in testicular cells (pre- and post- reductional germ cells, meiotic stages, spermatozoa and in Sertoli cells) from infertile individuals. 3) To analyze the frequency of chromosome abnormalities observed in the different stages of spermatogenesis and to compare these data with seminogram, meiotic studies and sperm FISH analysis in order to identify patients with a common pattern. MATHERIAL AND METHOD: Thirty-one diagnostic testicular biopsies from patients who had sought consultation for fertility problems have been provided for this study. By using the developed methodology, we have analyzed the presence of numerical chromosome abnormalities and apoptotic processes. In these samples analyzed we have evaluated the chromosome constitution of germ cells (pre- and post-reductional germ cells, meiotic figures and spermatozoa) and Sertoli cells in the three cell populations (apoptotic, viable and non-sorted). Therefore, taken together all patient data, we have analysed the differences between the percentages of chromosome abnormalities in the three populations of germ cells and Sertoli cells. Finally, based on the results obtained, a cluster analysis of patients was build. RESULTATS: The methodology is efficient to detect apoptotic processes and chromosomal abnormalities association in testicular cells. The analysis of the results has revealed a selection of aneuploid cells to the apoptotic fraction in pre-reductional germ cells and in spermatozoa. However, in Sertoli cells, meiotic figures and post-reductional germ cells no relationship between apoptosis and aneuploidy was observed. CONCLUSIONS: 1) The developed methodology allows the detection, isolation and cytogenetic analysis of apoptotic cells at different stages of spermatogenesis and in Sertoli cells from testicular biopsies. 2) Chromosome abnormalities observed in pre-reductional germ cells arise from mitotic segregation errors and are efficiently eliminated through apoptotic processes. 3) Primary spermatocytes are mostly euploid and no relationship between aneuploidies and apoptosis is observed. 4) Aneuploidy in Sertoli cells do not trigger cell death through apoptosis. 5) The aneuploidy level observed in post-meiotic germ cells demonstrates that the spindle assembly checkpoint (SAC) is not efficiently enough to eliminate aneuploid cells through apoptosis. No relationship between apoptosi and aneuploidy are observed in post-meiotic germ cells. These findings together with the observed reduction of aneuploid spermatozoa suggest that aneuploid post-meiotic germ cells are eliminated by apoptotic-independent cell death processes. 6) Aneuploid spermatozoa are depleted from the seminiferous tubules by apoptosis. 7) Our population is very heterogeneous and thus no similar clinical characteristics could be indentified in patients clusters according to the frequency of chromosome abnormalities at different stages of spermatogenesis. This is explained by the cytogenetic and phenotypic diversity of the infertile populations.

We designed a targeted-array called GOLD (Gain or Loss Detection) Chip consisting of 900 FISH-mapped non-overlapping BAC clones spanning the whole genome to enhance the coverage of 66 unique human genomic regions involved in well known microdeletion/microduplication syndromes. The array has a 10 Mb backbone to guarantee the detection of the aneuploidies, and has an implemented resolution for telomeres, and for regions involved in common genomic diseases. In order to evaluate clinical diagnostic applicability of GOLDChip, analytical validity was carried-out via retrospective analysis of DNA isolated from a series of cytogenetically normal amniocytes and cytogenetically abnormal DNA obtained from cultured amniocytes, peripheral blood and/or cell lines. We recruited 47 DNA samples corresponding to pathologies with significant frequencies (Cri du Chat syndrome, Williams syndrome, Prader Willi/Angelman syndromes, Smith-Magenis syndrome, DiGeorge syndrome, Miller-Dieker syndrome, chromosomes 13, 18 and 21 trisomies). We set up an experimental protocol that allowed to identify chromosomal rearrangements in all the DNA samples analyzed. Our results provide evidence that our targeted BAC array can be used for the identification of the most common microdeletion syndromes and common aneuploidies.

Objetivo: Describir los hallazgos ecográficos detectados en los fetos y su relación con el cariotipo anormal obtenido en las amniocentesis genéticas realizadas en el segundo trimestre. Métodos: Se revisaron todos los informes de los estudios ecográficos realizados previamente a la amniocentesis de los fetos que resultaron con cromosomopatías durante el año 2006. Ambiente: Centro Nacional de Genética Humana y Experimental. Universidad Central de Venezuela. Caracas Resultados: En 1786 amniocentesis, se obtuvieron 32 fetos (1,79 %) con cromosomopatías, siendo la trisomía 21 la más frecuente (47 %). De todos los casos con alteraciones cromosómicas 15 fetos (46,9 %) tuvieron uno o más marcadores ecográficos. Los hallazgos ecográficos encontrados en el grupo general fueron: pliegue nucal engrosado en 40 %, higroma quístico en 19,5 %, ectasia pielocalicial en 12,5 %, e intestino hiperecogénico, ausencia de hueso nasal, foco ecogénico intracardíaco y pie equino varo, cada uno en un 7 %. Un 53 % de los fetos con trisomía 21 presentaron algún tipo de hallazgo ecográfico anormal, así mismo un 67 % de los fetos con trisomía 18 y 100 % de los fetos que tenían monosomía X, trisomía 22 y triploidía Conclusión: El ultrasonido juega un papel muy importante en el diagnóstico prenatal. El diagnóstico apropiado de las anomalías estructurales y hallazgos menores, incrementa la sospecha de alguna alteración cromosómica y puede sugerir la realización de procedimientos invasivos de diagnóstico prenatal. Objective: To describe the abnormalities detected by detailed second trimester ultrasonography among the fetuses with abnormal karyotype after amniocentesis. Methods: Ultrasound studies belonging to fetuses with diagnosed chromosomal anomalies in 2006 were reviewed. Setting: Centro Nacional de Genética Humana y Experimental. Universidad Central de Venezuela. Caracas Results: A total of 1,786 patients underwent amniocentesis with a result of 32 (1.79 %) fetuses detected with an abnormal karyotype, trisomy 21 (47 %). Out of all cases with chromosomal anormalies 15 fetuses (46.9.%) had one or more sonographic markers. Ultrasound markers were: increased nucal fold 40 % , cystic hygroma (19.5 %) , pyelectasis 12.5 %, and echogenic bowel, absent nasal bone, echogenic intracardiac foci and club foot (7 %) each one. A 53 % of fetuses with trisomy 21 had some adnormal sonographic marker, as well as 67 % of trisomy 18 and 100 % of fetuses with X0, trisomy 22 and triploidy. Conclusions: The second trimester ultrasound plays an important role in prenatal diagnosis. The appropriate detection of structural anomalies and sonographic markers increase the diagnosis of aneuploidies and can suggest the necessity to practice an invasive study.

BACKGROUND: Patients with poor semen quality show increased sperm disomy and diploidy rates. Oligozoospermia and teratozoospermia are known to influence sperm aneuploidy, but there is still a debate about whether aneuploidies are associated with reduced motility. METHODS: Ejaculates from a large group of patients were examined by light microscopy to evaluate sperm concentration, motility and morphology, and by fluorescence in-situ hybridization (FISH) to analyse the presence of aneuploidies. Statistical analysis was performed to compare differences and to evaluate the relationship between sperm aneuploidy rate and semen quality. RESULTS: Five groups were established following the motility parameter, and total aneuploidy rates were statistically significantly higher in the groups where motility was

Los modelos actuales de carcinogenesis consideran la ocurrencia de mutaciones crecientes y mecanismos selectivos, favoreciendo la sobrevivencia y proliferacion celular aumentada. Mecanismos epigeneticos tambien participan en la oncogenesis, destacando la metilacion del DNA. La caracteristica de las celulas tumorales que permite el aumento de la ocurrencia de mutaciones es denominada inestabilidad genetica, donde son identificados dos mecanismos: inestabilidad de microsatelites, caracterizada por alteraciones nucleotidicas con errores en los sistemas de reparacion del DNA; inestabilidad cromosomica, en la cual las aberraciones suceden en grandes segmentos cromosomicos. Los carcinomas son caracterizados por alteraciones citogeneticas complejas y grandes mezclas genicas. Alteraciones telomericas, quiebres de DNA reparados inadecuadamente y deficiencia en los sistemas de chequeo del huso mitotico, son eventos capaces de generar inestabilidad cromosomica y aneuploidia que caracterizan estas neoplasias mas agresivas. El conocimiento de los mecanismos que provocan la inestabilidad cromosomica puede permitir la utilizacion clinica de informacion en el desarrollo de estrategias terapeuticas mas adecuadas, dirigidas a puntos especificos involucrados en procesos de malignizacion

In the current carcinogenesis models, the occurrence of increasing mutations and selection mechanisms favoring cell survival and higher proliferation rates are taken into account. Epigenetic mechanisms, among which DNA methylation stands out, also take part in oncogenesis. The characteristic of tumor cells that allows the increase of mutations is named genetic instability, encompassing two mechanisms: microsatellite instability, characterized by nucleotide alterations with errors in the DNA repair systems; and chromosomal instability, represented by aberrations occurring in large chromosome segments. Carcinomas are characterized by complex cytogenetic alterations and large gene amalgamations. Telomeric alterations, inadequately repaired DNA breaks, and deficiencies in the mitotic spindle checking systems are events capable of generating the chromosomal instability and aneuploidy which characterize more aggressive neoplasias. A better understanding of the chromosomal instability mechanisms can show the way towards a clinical utilization of such information, like developing more adequate therapeutic strategies, targeted at specific sites involved in the malignization process Los modelos actuales de carcinogénesis consideran la ocurrencia de mutaciones crecientes y mecanismos selectivos, favoreciendo la sobrevivencia y proliferación celular aumentada. Mecanismos epigenéticos también participan en la oncogénesis, destacando la metilación del DNA. La característica de las células tumorales que permite el aumento de la ocurrencia de mutaciones es denominada inestabilidad genética, donde son identificados dos mecanismos: inestabilidad de microsatélites, caracterizada por alteraciones nucleotídicas con errores en los sistemas de reparación del DNA; inestabilidad cromosómica, en la cual las aberraciones suceden en grandes segmentos cromosómicos. Los carcinomas son caracterizados por alteraciones citogenéticas complejas y grandes mezclas génicas. Alteraciones teloméricas, quiebres de DNA reparados inadecuadamente y deficiencia en los sistemas de chequeo del huso mitótico, son eventos capaces de generar inestabilidad cromosómica y aneuploidía que caracterizan estas neoplasias más agresivas. El conocimiento de los mecanismos que provocan la inestabilidad cromosómica puede permitir la utilización clínica de información en el desarrollo de estrategias terapéuticas más adecuadas, dirigidas a puntos específicos involucrados en procesos de malignización

Pericentric inversions involving the secondary constriction (qh) region of chromosome 9 are considered to be normal variants of human karyotype. A number of investigators have suggested that chromosomal anomalies can contribute to human infertility causing spermatogenetic derangement. The present study was aimed at verifying the influence of chromosome 9 inversion on human spermatogenesis. Semen samples of 18 male carriers of chromosome 9 inversion, analysed by light microscopy, revealed that five patients were azoospermic. PCR analysis demonstrated that two of them also had Y microdeletions. The other 13 showed generally normal sperm concentrations and reduced motility. The morphological characteristics of sperm were studied by TEM and the data were elaborated by a mathematical formula. Sperm pathologies resulted more frequently in the studied group compared to controls, particularly apoptosis. Partial sequences of the A-kinase anchoring protein (Akap) 4 and 3 genes were performed in all patients, as a previous study by our group highlighted Dysplasia of Fibrous Sheath (DFS) defect in two men with inv 9 investigations. The possible effect of chromosome 9 inversion on meiotic chromosome segregation was investigated by FISH, which showed an increased incidence of diploidy. We hypothesized that this inversion could have variable effects on spermatogenesis, from azoospermia to severely altered sperm morphology, motility and meiotic segregation.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:21:48Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:40:03Z : No. of bitstreams: 1 2-s2.0-33644907463.pdf: 1009529 bytes, checksum: ae9f5a1a572900a8f00e24445d9eb858 (MD5) Made available in DSpace on 2014-05-27T11:21:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-01-28 Aim: To investigate the occurrence of chromosome 3, 7, 8, 9, and 17 aneuploidies, TP53 gene deletion and p53 protein expression in chronic gastritis, atrophic gastritis and gastric ulcer, and their association with H pylori infection. Methods: Gastric biopsies from normal mucosa (NM, n = 10), chronic gastritis (CG, n = 38), atrophic gastritis (CAG, n = 13) and gastric ulcer (GU, n = 21) were studied using fluorescence in situ hybridization (FISH) and immunohistochemical assay. A modified Giemsa staining technique and PCR were used to detect H pylori. An association of the gastric pathologies and aneuploidies with H pylori infection was assessed. Results: Aneuploidies were increasingly found from CG (21%) to CAG (31%) and to GU (62%), involving mainly monosomy and trisomy 7, trisomies 7 and 8, and trisomies 7, 8 and 17, respectively. A significant association was found between H pylori infection and aneuploidies in CAG (P = 0.0143) and GU (P = 0.0498). No TP53 deletion was found in these gastric lesions, but p53-positive immunoreactivity was detected in 45% (5/11) and 12% (2/17) of CG and GU cases, respectively. However, there was no significant association between p53 expression and H pylori infection. Conclusion: The occurrence of aneuploidies in benign lesions evidences chromosomal instability in early stages of gastric carcinogenesis associated with H pylori infection, which may confer proliferative advantage. The increase of p53 protein expression in CG and GU may be due to overproduction of the wild-type protein related to an inflammatory response in mucosa. © 2006 The WJG Press. All rights reserved. UNESP - São Paulo State University, Rua Cristovao Colombo 2265, 15054-000 Sao Jose do Rio Preto, SP UNIP - Araçatuba, Araçatuba, SP Faculdades Católicas Salesianas, Araçatuba, SP UNESP - São Paulo State University Department of Biology, Rua Cristovao Colombo 2265, 15054-000 Sao Jose do Rio Preto FAMERP School of Medicine, Sao Jose do Rio Preto, SP UNESP - São Paulo State University, Rua Cristovao Colombo 2265, 15054-000 Sao Jose do Rio Preto, SP UNESP - São Paulo State University Department of Biology, Rua Cristovao Colombo 2265, 15054-000 Sao Jose do Rio Preto

Cytogenetic results of amniocentesis materials: incidence of abnormal karyotypes in the Turkish collaborative study: The experience on prenatal chromosome diagnosis of four Turkish centers participating in a collaborative study on 6041 genetic amniocentesis performed during a 4-8 years period were reviewed. 5887 (97.5%) patients had strong clinical indications for prenatal chromosome studies and 154 (2.5%) were referred because of maternal anxiety and a bad history of previous gestations. The main indication groups were: advanced maternal age (3197 cases), positive serum screening (2011 cases), ultrasound-identified anomaly (492 cases), previous fetus/child with chromosomal aberrations (103 cases), a history of a previous abnormal and / or mentally handicapped child (70 cases) and a parental chromosome rearrangement (14 cases). The average maternal age was 33.9 years and average gestational age was 18 weeks. A total of 179 affected fetuses were detected in this collaborative study (3%) of which 133 were unbalanced (74.3%). Among the 124 (69%) numerical aberrations, 102 (82.3%) were autosomal aneuploidies, 20 (16.1%) were gonosomal aneuploidies and 2 (1.6%) were poliploidies. Among the 55 (31%) structural aberrations, balanced translocation was the most common (63.6%) and I 1 cases of inversion, four cases of unbalanced translocation, two cases of marker chromosome and three cases of other abnormalities were found. The overall culture success rate was 99.7%. Pregnancy termination that is permitted by legal authorities was accepted by 94.7% (126/133) with parents at unbalanced cytogenetic result announcement.

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The isolation and analysis of nucleated fetal cells (NFCs) from maternal blood may represent a new approach to noninvasive prenatal diagnosis. Although promising, these techniques require highly accurate separation of NFCs from nucleated cells of maternal origin; the two major problems limiting these techniques are the relative rarity of fetal cells in maternal blood and the need to establish their fetal origin. We now report a novel procedure that has allowed accurate separation of NFCs from maternal cells. The technique reported involves direct micromanipulator isolation of histochemically identified hemoglobin F-positive nucleated cells to obtain fetal nucleated red blood cells (FNRBCs) of high yield and purity. Using this technique, followed by cell-by-cell multicolor fluorescence in situ hybridization (FISH) analysis of purified FNRBCs, we were able to detect some of the most common human aneuploidies (including Down syndrome, Klinefelter syndrome, and trisomy 13) in 33 pregnant women referred for amniocentesis. The procedure used, which can be completed in

Currently, prenatal detection of Down syndrome and other most common aneuploidies relies on invasive procedures such as amniocentesis and villocentesis, and on non-invasive screening tests such as second trimester maternal serum screening (Triple test), and first trimester screening (ULTRA-screen). However, it well known that invasive techniques carry a small risk of fetal loss, while both Triple test and ULTRA-screen are not diagnostic, may miss from 15 - 40 % of cases of Down syndrome, in addition to having a 5 - 8 % rate of false-positives. We now report clear evidence that the number of fetal nucleated red blood cells (FNRBCs) in the maternal circulation is remarkably higher in pregnant women carrying aneuploid fetuses, especially in cases of Down syndrome. These results are in agreement with the findings of other investigators using different methods, and suggest that the number of FNRBCs present in the maternal blood sample could be used as additional marker, in concert with existing screening tests, to improve non-invasive detection of Down syndrome, and other most common aneuploidies.

Aims and Objective: Primed in situ labeling/synthesis (PRINS) technique is an alternative to fluorescent in situ hybridization for chromosome analysis. This study was designed to evaluate the application of PRINS for rapid diagnosis of common chromosomal aneuploidy. Materials and Methods : We have carried out PRINS using centromere specific oligonucleotide primers for chromosome X, Y, 13, 18 and 21 on lymphocyte metaphase and interphase cells spread. Specific primer was annealed in situ , followed by elongation of primer by Taq DNA polymerase in presence of labeled nucleotides. Finally, reaction was stopped and visualized directly under fluorescent microscope. Results: Discrete centromere specific signals were observed with each primer. Conclusion: PRINS seems to be a rapid and reliable method to detect common chromosome aneuploidy in peripheral blood lymphocyte metaphase and interphase cells.

The clinical application of preimplantation genetic diagnosis (PGD) for aneuploidy has confirmed the hypothesis that implantation failure and spontaneous abortions are frequently due to aneuploidy. Following PGD, a higher implantation rate and a lower incidence of spontaneous abortions are obtained in patient categories where aneuploidy is the main cause of reproductive failure: women in advanced reproductive age, patients with an altered karyotype due to translocations or gonosomal mosaicism, and patients with recurrent spontaneous abortions. In these cases, the transfer of euploid embryos overcomes the poor prognosis condition in these couples. As expected, aneuploidy increases proportionally with female age; however, not all the chromosomes studied show this trend, suggesting that segregation errors could occur at different rates for each chromosome in relation to maternal age. Furthermore, the retrospective analysis of the results obtained in patients who repeated at least twice a PGD cycle permitted to estimate their chances of reproducing the same pattern of chromosomal abnormalities and consequently evaluating their possibility of a pregnancy: when no euploid embryos are detected at the first attempt, the chance of on-term pregnancy is below 10%; however, this chance is approximately 30% for couples with at least two euploid embryos in the first cycle.