Fetal exome sequencing: yield and limitations in a tertiary referral center.

OBJECTIVE: To explore the indications for and diagnostic outcomes of fetal exome sequencing in a tertiary referral center. METHODS: Between 2012 and 2017, 77 unrelated fetal samples from pregnancies referred to our center underwent exome sequencing. The cohort included 37 fetuses, 36 products of conception (from cases of pregnancy termination or intrauterine fetal death), one case with DNA from both the fetus and a previous termination of pregnancy, and three cases with DNA of unknown origin. Exome sequencing was performed on DNA extracted from amniocytes or fetal tissue and, in some cases, from parental peripheral blood. Indications, turnaround time, diagnostic rates and pregnancy outcomes were investigated. Diagnostic yield was analyzed according to consanguinity (yes or no), sample type (proband only, or trio or other) and referral indication (malformation or isolated nuchal translucency (NT)). RESULTS: The most common indication for fetal exome sequencing was multiple malformations (21/77, 27%), followed by isolated brain malformation (15/77, 19%). Twelve (16%) fetuses were referred for isolated increased NT. Exome analysis was diagnostic for 16 fetuses (21%); when subclassified into fetal malformations vs isolated increased NT it became clear that exome analysis did not reveal any known or probable pathogenic variants in cases referred for isolated increased NT, whereas, among the remaining fetuses, a molecular diagnosis was reached in 16/65 (25%). Proband-only cases received a diagnosis more often than did cases that had trio exome sequencing. CONCLUSIONS: Exome sequencing has the potential to provide molecular diagnoses in cases in which conventional prenatal cytogenetic testing is negative. Referral bias of consanguineous cases could account for the high diagnostic rate of proband-only sequencing. Syndrome-specific prognostic information enables parents to make informed decisions, whereas challenges include time limitations and variant interpretation in the setting of non-specific fetal findings. As we report only established disease-gene associations, further segregation and functional studies in a research setting are expected to increase significantly the diagnostic yield. Copyright © 2018 ISUOG. Published by John Wiley & Sons Ltd.

The interaction between neurocognitive functioning, subthreshold psychotic symptoms and pharmacotherapy in 22q11.2 deletion syndrome: A longitudinal comparative study.

BACKGROUND: The 22q11.2 deletion syndrome (22q11DS) is the most common genetic syndrome associated with schizophrenia. The goal of this study was to evaluate longitudinally the interaction between neurocognitive functioning, the presence of subthreshold psychotic symptoms (SPS) and conversion to psychosis in individuals with 22q11DS. In addition, we attempted to identify the specific neurocognitive domains that predict the longitudinal evolution of positive and negative SPS, as well as the effect of psychiatric medications on 22q11DS psychiatric and cognitive developmental trajectories. METHODS: Forty-four participants with 22q11DS, 19 with Williams syndrome (WS) and 30 typically developing (TD) controls, age range 12-35years, were assessed at two time points (15.2±2.1months apart). Evaluation included the Structured Interview for Prodromal Symptoms (SIPS), structured psychiatric evaluation and the Penn Computerized Neurocognitive Battery (CNB). RESULTS: 22q11DS individuals with SPS had a yearly conversion rate to psychotic disorders of 8.8%, compared to none in both WS and TD controls. Baseline levels of negative SPS were associated with global neurocognitive performance (GNP), executive function and social cognition deficits, in individuals with 22q11DS, but not in WS. Deficits in GNP predicted negative SPS in 22q11DS and the emergence or persistence of negative SPS. 22q11DS individuals treated with psychiatric medications showed significant improvement in GNP score between baseline and follow-up assessments, an improvement that was not seen in untreated 22q11DS. CONCLUSIONS: Our results highlight the time-dependent interplay among positive and negative SPS symptoms, neurocognition and pharmacotherapy in the prediction of the evolution of psychosis in 22q11DS.

Genomic disorders 20 years on-mechanisms for clinical manifestations.

Genomic disorders result from copy-number variants (CNVs) or submicroscopic rearrangements of the genome rather than from single nucleotide variants (SNVs). Diverse technologies, including array comparative genomic hybridization (aCGH) and single nucleotide polymorphism (SNP) microarrays, and more recently, whole genome sequencing and whole-exome sequencing, have enabled robust genome-wide unbiased detection of CNVs in affected individuals and in reportedly healthy controls. Sequencing of breakpoint junctions has allowed for elucidation of upstream mechanisms leading to genomic instability and resultant structural variation, whereas studies of the association between CNVs and specific diseases or susceptibility to morbid traits have enhanced our understanding of the downstream effects. In this review, we discuss the hallmarks of genomic disorders as they were defined during the first decade of the field, including genomic instability and the mechanism for rearrangement defined as nonallelic homologous recombination (NAHR); recurrent vs nonrecurrent rearrangements; and gene dosage sensitivity. Moreover, we highlight the exciting advances of the second decade of this field, including a deeper understanding of genomic instability and the mechanisms underlying complex rearrangements, mechanisms for constitutional and somatic chromosomal rearrangements, structural intra-species polymorphisms and susceptibility to NAHR, the role of CNVs in the context of genome-wide copy number and single nucleotide variation, and the contribution of noncoding CNVs to human disease.

Charcot-Marie-Tooth disease and pathways to molecular based therapies.

The discovery in 1991 that chromosome 17p12 duplication is associated with Charcot-Marie-Tooth (CMT) disease marked the beginning of an era of molecular insight into this disorder, which encompasses the peripheral motor and sensory neuropathies. A mere two decades later, over 40 subtypes of CMT have been molecularly defined and many have been extensively studied in vitro and in animal models, providing the framework for a more comprehensive understanding of the biological pathways dictating myelination, axonal dynamics, and axon-glia interactions. The advent of next-generation sequencing technologies offers opportunities in both research and clinical settings for gene discovery, further molecular understanding and diagnosis, and calls for modifications of the existing algorithms guiding genetic testing. Although treatment is mainly supportive at this time, advances in this field are anticipated as the molecular basis of CMT is unraveled.

Co-morbidity of Emery-Dreifuss muscular dystrophy and a congenital myasthenic syndrome possibly affecting the phenotype in a large Bedouin kindred.

Emery-Dreifuss muscular dystrophy (EDMD) is an X-linked humero-peroneal muscular dystrophy associated with contractures and cardiomyopathy. In a 90 member family, we found 11 affected male individuals, three of whom displayed areflexia and neurogenic electromyographic changes. Muscle biopsy performed in one case demonstrated type grouping suggestive of a neurogenic disorder. These three individuals and another family member, who suffers from mild, static limb weakness but is clinically and genetically unaffected by EDMD showed an abnormal incremental response of over 100% to tetanic stimulation. In contrast, one affected family member showed myopathic features on needle electromyography and no definite pathology in repetitive stimulation studies. The diagnosis of EDMD was established by demonstrating a 1712_1713insTGGGC mutation in the emerin gene. This family apparently expresses co-morbidity of EDMD with an exceptionally mild form of pre-synaptic congenital myasthenic syndrome resembling the Lambert-Eaton myasthenic syndrome (LEMS). The superimposed pre-synaptic disorder may have contributed to the development of the neurogenic features demonstrated in these patients.

Mitochondrial metabolism reveals a functional architecture in intact islets of Langerhans from normal and diabetic Psammomys obesus.

The cells within the intact islet of Langerhans function as a metabolic syncytium, secreting insulin in a coordinated and oscillatory manner in response to external fuel. With increased glucose, the oscillatory amplitude is enhanced, leading to the hypothesis that cells within the islet are secreting with greater synchronization. Consequently, non-insulin-dependent diabetes mellitus (NIDDM; type 2 diabetes)-induced irregularities in insulin secretion oscillations may be attributed to decreased intercellular coordination. The purpose of the present study was to determine whether the degree of metabolic coordination within the intact islet was enhanced by increased glucose and compromised by NIDDM. Experiments were performed with isolated islets from normal and diabetic Psammomys obesus. Using confocal microscopy and the mitochondrial potentiometric dye rhodamine 123, we measured mitochondrial membrane potential oscillations in individual cells within intact islets. When mitochondrial membrane potential was averaged from all the cells in a single islet, the resultant waveform demonstrated clear sinusoidal oscillations. Cells within islets were heterogeneous in terms of cellular synchronicity (similarity in phase and period), sinusoidal regularity, and frequency of oscillation. Cells within normal islets oscillated with greater synchronicity compared with cells within diabetic islets. The range of oscillatory frequencies was unchanged by glucose or diabetes. Cells within diabetic (but not normal) islets increased oscillatory regularity in response to glucose. These data support the hypothesis that glucose enhances metabolic coupling in normal islets and that the dampening of oscillatory insulin secretion in NIDDM may result from disrupted metabolic coupling.

System response of the sinoatrial node during vagal stimulation.

The SA node response to modulations in canine vagal tone was investigated by means of the heart rate variability power spectrum. A new algorithm that was developed for accurate power spectrum estimation of short R-R segments is described. The performance of the algorithm was assessed for ECG recordings obtained from a controlled experiment, in which a frequency modulated pulse train was applied to the vagal nerve after vagal transaction and blockade of the sympathetic system. The power spectrum calculated for 20-25 heartbeats showed conspicuous spectral peaks in accordance with the different modulating frequencies between 0.1 and 0.3 Hz. The presence of spurious peaks was negligible even when the analysed signal segment consisted of only 20-25 beats. These results imply that for a certain range of modulating frequencies the sinoatrial node responds linearly to fluctuations in the parasympathetic tone. System identification methods that include fitting a linear model to the heart rate variability signal and analysis of the residuals were used for confirming the hypothesis of linearity. For higher frequencies of the modulating signal, usually above 0.3 Hz, the system was found to deviate from linearity.

Altered heart rate variability in panic disorder patients.

Resting electrocardiographic recordings were obtained from 10 patients with panic disorder (PD) and 14 normal controls. Signal analysis of the beat-to-beat heart rate variability was performed by means of power spectrum analysis. The analysis revealed that patients with PD had marked reduction in the high-frequency peaks of the power spectrum densities. An Energy Ratio Index (ERI), which accurately differentiated between patients and controls, was calculated. In PD patients, a significant correlation was demonstrated between the clinical ratings and the energy ratios. Our findings suggest that decreased heart rate variability may be a characteristic of PD. The importance of this finding as a diagnostic marker and the underlying pathophysiological mechanisms need to be further explored.

Increased sympathetic and decreased parasympathetic cardiac innervation in patients with Alzheimer's disease.

To assess whether Alzheimer's disease affects the sympathetic and parasympathetic influences on the heart rate, we used power spectrum analysis of heart rate variability derived from simple body-surface electrocardiography. We calculated the energy ratio of low- to high-frequency bands. This ratio was significantly higher in patients with Alzheimer's disease than in normal controls (upright posture, 0.41 +/- 0.21 vs 0.23 +/- 0.08). The parasympathetically mediated baroreceptor activity reflected by the energy ratio of medium- to low- and high-frequency bands was significantly depressed in patients with Alzheimer's disease (upright posture, 0.12 +/- 0.02 vs 0.07 +/- 0.03; supine posture, 0.11 +/- 0.02 vs 0.085 +/- 0.025). Compared with normal volunteer controls, patients with Alzheimer's disease manifested a relatively hypersympathetic, hypoparasympathetic state.

Retinal detachment with high myopia in the Rubinstein-Taybi syndrome.

A case of rhegmatogenous retinal detachment with high myopia is presented in a 17 year old boy with the typical characteristics of the Rubinstein-Taybi syndrome. Although multiple eye anomalies are known to occur in this syndrome, the occurrence of retinal detachment has not been reported up to now. The importance of including a thorough fundus examination in the routine eye examination of these patients is emphasized.

Enhancement of the response of ageing mouse lymphocytes by in vitro treatment with lecithin.

The effect of treatment with phosphatidylcholine (lecithin) has been examined on splenocytes from young (6 months) and aged (greater than 20 months) (C3H/eB X C57BL/6)F1 male mice, by probing their responsiveness to proliferative signals of mitogens and mixed lymphocyte reaction. This study was initiated since old mice have been known to manifest an increased ratio of cholesterol: phospholipids (C/PL) in their plasma membranes. Unlike the case in young mice, enhancement in proliferative responses was demonstrated on cells from the old after incubation with lecithin. This enhancement was achieved using a variety of methods by which lymphocytes were exposed to the lipids, and which were all designed to reduce the C/PL ratio in the plasma membrane. The observed differences in response to lecithin treatment between young and old did not stem from different extents of lipid incorporation, and seems unlikely to be a result of modified binding of the mitogens after treatment with the lipid. The data suggest that the normal functional performance of specific reactions, which decline with age, may be restored artificially at an appropriate age, if other complementary functions have not been damaged.

The relevance of ideas and beliefs to immigration as it is seen from a theory of personality point of view.

In the realm of human culture, ideas and beliefs are studied in the disciplines of sociology, philosophy, not psychology. Although human spiritual abilities, that is, the ability to speak, think, yield thoughts, ideas, beliefs are maybe that which best characterize the human being as such, they are not considered part of the study of human personality. The levels-of-organization model of personality, which is presented here, considers human spiritual powers as part and parcel of human personality, and ascribes to it a very important specialized role. According to it, the highest stage of human growth are his spiritual abilities. Consequently, whatever a person does depends a great deal on what he thinks and believes. The importance of human spiritual powers is best manifested in crisis situations. This presentation will consider immigration as an example to stress, and examine the contribution of human ideas and beliefs to human ability to overcome this kind of stress.