Claudio Giorlandino

Oxidative stress occurs early in Down syndrome pregnancy: A redox proteomics analysis of amniotic fluid

Purpose: The present study aims to evaluate a set of oxidative stress biomarkers in the amniotic fluid (AF) of women carrying Down syndrome (DS) fetuses that could prove in vivo the early occurrence of oxidative damage in DS.

Spermatozoa with chromosomal abnormalities may result in a higher rate of recurrent abortion

OBJECTIVE To determine whether sperm aneuploidy can lead to abortion. DESIGN Clinical study. SETTING Couples with reproductive problems evaluated in a private diagnostic laboratory. PATIENT(S) Two men whose wives had histories of multiple abortions. INTERVENTION(S) Whole and Percoll-processed semen samples were analyzed. MAIN OUTCOME MEASURE(S) The results of fluorescence in situ hybridization. RESULT(S) Aneuploidy rates in Percoll-processed samples were higher than those found in whole specimens. Aneuploid spermatozoa also displayed greater motility. CONCLUSION(S) Sperm aneuploidy should be studied before and after Percoll capacitation in all couples with unexplained infertility.

BLOOD CONTAMINATION OF AMNIOTIC FLUID AFTER AMNIOCENTESIS IN RELATION TO PLACENTAL LOCATION

The aim of the present study was to evaluate blood contamination of the amniotic fluid collected in 20 patients undergoing a second amniocentesis performed 2 weeks after a first procedure that had failed due to Pseudomonas aeruginosa contamination of the cell cultures. Red blood cell and haemoglobin concentrations in the amniotic fluid were significantly higher in patients who had undergone a transplacental procedure compared with patients in whom the placenta was not traversed with the needle. For both groups, blood contamination of the amniotic fluid was significantly higher compared with a control group of 20 patients undergoing amniocentesis for the first time. Significant blood contamination of the amniotic fluid after amniocentesis occurs in every instance if evaluated at a ‘second‐look’ procedure; the blood contamination is higher when an anterior placenta is traversed with the needle. The clinical significance of these findings needs to be further evaluated.

Antibiotic Prophylaxis before second-trimester Genetic Amniocentesis (APGA): a single-centre open randomised controlled trial

To compare procedure‐related pregnancy loss after second‐trimester genetic amniocentesis in women given an antibiotic prophylaxis and controls.

The role of ultrasonography in the diagnosis of fetal isolated complete agenesis of the corpus callosum: a long-term prospective study.

Objective. To evaluate the role of a dedicated neurosonographer in prenatal diagnosis of isolated complete agenesis of the corpus callosum (iCACC) and to asses the postnatal outcome of these infants. Methods. Prospective study between January 2004 to December 2004 at Fetal Maternal Medical Centre ‘Artemisia’, Rome, Italy. A detailed ultrasound scan was performed in fetuses affected by iCACC by a dedicated fetal neurosonographer (CG). In all cases, magnetic resonance imaging (MRI) within 5 weeks and 13–15 months after birth was performed. A comparison was made between prenatal findings following the ultrasound scan and postnatal MRI. In these cases, a follow-up of 4-years was performed with a neurological evaluation. Results. Among 23 cases of ACC diagnosed at our centre in the study period, CACC was diagnosed in 17 fetuses. Two were then excluded due to associated malformations, one was lost at follow-up and one patient opted to terminate her pregnancy. Newborn MRI confirmed the ultrasonographic diagnosis of iCACC in all 13 cases. A regular development was present in 92.3% of prenatally diagnosed iCACC. Conclusion. A dedicated neurosonographer could diagnose the iCACC with the same accuracy as MRI and in up to 90% of cases the newborn will have a regular development.

Reference charts for fetal corpus callosum length: A prospective cross-sectional study of 2950 fetuses

The purpose of this study was to establish reference charts for fetal corpus callosum length in a convenience sample.

What Is the Rate of Incomplete Fetal Anatomic Surveys During a Second-Trimester Scan? Retrospective Observational Study of 4000 Nonobese Pregnant Women

The purpose of this study was to estimate the rate of incomplete fetal anatomic surveys during a second‐trimester scan due to an unfavorable fetal position in a nonobese population.

Pallister–Killian syndrome: Cytogenetics and molecular investigations of mosaic tetrasomy 12p in prenatal chorionic villus and in amniocytes. Strategy of prenatal diagnosis

OBJECTIVE Pallister-Killian syndrome (PKS) is a rare, sporadic genetic disorder caused by mosaic tetrasomy of the short arm of chromosome 12 (12p). Clinically, PKS is characterized by several systemic abnormalities, such as intellectual impairment, hearing loss, epilepsy, hypotonia, craniofacial dysmorphism, pigmentary skin anomalies, epilepsy, and a variety of congenital malformations. Prenatally, PKS can be suspected in the presence of ultrasound anomalies: diaphragmatic hernia, rhizomelic micromelia, hydrops fetalis, fetal overweight, ventriculomegaly in the central nervous system, congenital heart defects, or absent visualization of the stomach. In all these cases, a detailed genetic study is required. PKS is diagnosed by prenatal genetic analysis through chorionic villus sampling, genetic amniocentesis, and cordocentesis. CASE REPORT We report two cases of PKS with prenatal diagnosis of isochromosome 12p made by cytogenetic studies. The first case is of a 36-year-old pregnant woman who underwent genetic chorionic villus sampling at 13th weeks of gestation after 1st trimester prenatal ultrasound revealed clinical features of PKS: flat nasal bridge and fetal hydrops. The second case is of a 32-year-old pregnant woman with genetic amniocentesis at 17th weeks of gestation that showed mos46,XX[21]/47,XX,+i(12p) associated to PKS. CONCLUSION New molecular cytogenetic techniques array comparative genomic hybridization and fluorescence in-situ hybridization in association with conventional karyotype are pivotal innovative tools to search for chromosomic anomalies and for a complete prenatal diagnosis, especially in cases such as PKS where array comparative genomic hybridization analysis alone could not show mosaicism of i(12p).

Comparative Study of a CGH and Next Generation Sequencing (NGS) forChromosomal Microdeletion and Microduplication Screening

BACKGROUND prenatal genetic diagnosis of rare disorders is undergoing in recent years a significant enhancement through the application of methods of massive parallel sequencing. Despite the quantity and quality of the data produced, just few analytical tools and software have been developed in order to identify structural and numerical chromosomal anomalies through NGS, mostly not compatible with benchtop NGS platform and routine clinical diagnosis. METHODS we developed technical, bioinformatic, interpretive and validation pipelines for Next Generation Sequencing to identify SNPs, indels, aneuploidies, and CNVs (Copy Number Variations). RESULTS we show a new targeted resequencing approach applied to prenatal diagnosis. For sample processing we used an enrichment method for 4,813 genes library preparation; after sequencing our bioinformatic pipelines allowed both SNPs analysis for approximately thirty diseases or diseases family involved in fetus development and numerical chromosomal anomalies screening. CONCLUSIONS results obtained are compatible with those obtained through the gold standard technique, aCGH array, moreover allowing identification of genes involved in chromosome deletions or duplications and exclusion of point mutation on allele not affected by chromosome aberrations.

Laser-assisted hatching of human embryos: may two alternative approaches (thinning versus drilling) impact on implant rate?

Oocytes and early embryos are surrounded by a 13–15-μm thick of zona pellucida layer that is involved in the formation of binding with the spermatozoa, hardening process, induction of acrosome reaction, egg fusion, and prevention of polyspermy [1]. Its function becomes less important once the structural junctions between blastomeres in the compaction process are established [2]. Hatching is the physiological process in which the embryo, at the blastocyst stage, comes out of the zona pellucida. This process is a fundamental prerequisite for implantation [3]. Several studies indicate that a zona pellucida thinning is an active process of embryo development already in the early stages [4], and it was demonstrated that not-cleaved embryos do not show changes in thickness of the zona pellucida in culture [5]. The inability of the blastocyst to hatch from the zona pellucida may be one of the factors involved in the high implant failure rate in human in vitro fertilization (IVF) [6], maybe due to a loss of elasticity of the zona pellucida in IVF embryos, similarly to an aging process [7] and to a reduced sensitivity to low pH. Assisted hatching (AH) is the methodology used in IVF to mimic the normal hatching stage of embryonic development and it should increase the implant rate among poor prognosis patients or on embryos noted to have a thick zona pellucida [8, 9]. It has been suggested that AH facilitates the implantation because it allows an early contact between embryo and endometrial tissues and exposing it to growth factors fundamental for the subsequent stages of the blastocyst’s development [10]. AH can be obtained by different methods: mechanical, chemical, or by laser energy. The mechanical techniques are performed by micromanipulator stabilizing the embryo by a holding pipette while a microneedle is pushed tangentially through the zona pellucida. This technique is quick to perform but the size of the hole cannot be standardized. The chemical approach is performed by preloading the microneedle with acid Tyrode and injecting it in a blastomere-free area. This technique requires a very high skill in handling the micromanipulator to perform it as quick as possible to prevent an unnecessary exposure of the embryo to the toxic acid solution. Laser-assisted hatching (LAH) can be applied through different techniques: drilling that is an artificial hole of the zona pellucida, just to the opposite side where the embryo is formed, and thinning that is just a thinning of the outer protective glycoprotein layer. LAH has the benefit of a non-contact mode on the embryos and is the easiest approach to perform. However, although many works compare the various methods [11], there are insufficient data that compare the different techniques within the same method [12]. The primary aim of this work is to retrospectively compare the implant rate after the application of two different techniques (drilling and thinning) obtained by LAH. * Stella Capriglione s.capriglione@unicampus.it