CFTR function is impaired in a subset of patients with pancreatitis carrying rare CFTR variants.

BACKGROUND: Many affected by pancreatitis harbor rare variants of the cystic fibrosis (CF) gene, CFTR, which encodes an epithelial chloride/bicarbonate channel. We investigated CFTR function and the effect of CFTR modulator drugs in pancreatitis patients carrying CFTR variants. METHODS: Next-generation sequencing was performed to identify CFTR variants. Sweat tests and nasal potential difference (NPD) assays were performed to assess CFTR function in vivo. Intestinal current measurement (ICM) was performed on rectal biopsies. Patient-derived intestinal epithelial monolayers were used to evaluate chloride and bicarbonate transport and the effects of a CFTR modulator combination: elexacaftor, tezacaftor and ivacaftor (ETI). RESULTS: Of 32 pancreatitis patients carrying CFTR variants, three had CF-causing mutations on both alleles and yielded CF-typical sweat test, NPD and ICM results. Fourteen subjects showed a more modest elevation in sweat chloride levels, including three that were provisionally diagnosed with CF. ICM indicated impaired CFTR function in nine out of 17 non-CF subjects tested. This group of nine included five carrying a wild type CFTR allele. In epithelial monolayers, a reduction in CFTR-dependent chloride transport was found in six out of 14 subjects tested, whereas bicarbonate secretion was reduced in only one individual. In epithelial monolayers of four of these six subjects, ETI improved CFTR function. CONCLUSIONS: CFTR function is impaired in a subset of pancreatitis patients carrying CFTR variants. Mutations outside the CFTR locus may contribute to the anion transport defect. Bioassays on patient-derived intestinal tissue and organoids can be used to detect such defects and to assess the effect of CFTR modulators.

Hybrid Shell-Beam Inverse Finite Element Method for the Shape Sensing of Stiffened Thin-Walled Structures: Formulation and Experimental Validation on a Composite Wing-Shaped Panel.

This work presents a novel methodology for the accurate and efficient elastic deformation reconstruction of thin-walled and stiffened structures from discrete strains. It builds on the inverse finite element method (iFEM), a variationally-based shape-sensing approach that reconstructs structural displacements by matching a set of analytical and experimental strains in a least-squares sense. As iFEM employs the finite element framework to discretize the structural domain and as the displacements and strains are approximated using element shape functions, the kind of element used influences the accuracy and efficiency of the iFEM analysis. This problem is addressed in the present work through a novel discretization scheme that combines beam and shell inverse elements to develop an iFEM model of the structure. Such a hybrid discretization paradigm paves the way for more accurate shape-sensing of geometrically complex structures using fewer sensor measurements and lower computational effort than traditional approaches. The hybrid iFEM is experimentally demonstrated in this work for the shape sensing of bending and torsional deformations of a composite stiffened wing panel instrumented with strain rosettes and fiber-optic sensors. The experimental results are accurate, robust, and computationally efficient, demonstrating the potential of this hybrid scheme for developing an efficient digital twin for online structural monitoring and control.

Non-Destructive Techniques for the Condition and Structural Health Monitoring of Wind Turbines: A Literature Review of the Last 20 Years.

A complete surveillance strategy for wind turbines requires both the condition monitoring (CM) of their mechanical components and the structural health monitoring (SHM) of their load-bearing structural elements (foundations, tower, and blades). Therefore, it spans both the civil and mechanical engineering fields. Several traditional and advanced non-destructive techniques (NDTs) have been proposed for both areas of application throughout the last years. These include visual inspection (VI), acoustic emissions (AEs), ultrasonic testing (UT), infrared thermography (IRT), radiographic testing (RT), electromagnetic testing (ET), oil monitoring, and many other methods. These NDTs can be performed by human personnel, robots, or unmanned aerial vehicles (UAVs); they can also be applied both for isolated wind turbines or systematically for whole onshore or offshore wind farms. These non-destructive approaches have been extensively reviewed here; more than 300 scientific articles, technical reports, and other documents are included in this review, encompassing all the main aspects of these survey strategies. Particular attention was dedicated to the latest developments in the last two decades (2000-2021). Highly influential research works, which received major attention from the scientific community, are highlighted and commented upon. Furthermore, for each strategy, a selection of relevant applications is reported by way of example, including newer and less developed strategies as well.

A Comparative Analysis of Signal Decomposition Techniques for Structural Health Monitoring on an Experimental Benchmark.

Signal Processing is, arguably, the fundamental enabling technology for vibration-based Structural Health Monitoring (SHM), which includes damage detection and more advanced tasks. However, the investigation of real-life vibration measurements is quite compelling. For a better understanding of its dynamic behaviour, a multi-degree-of-freedom system should be efficiently decomposed into its independent components. However, the target structure may be affected by (damage-related or not) nonlinearities, which appear as noise-like distortions in its vibrational response. This response can be nonstationary as well and thus requires a time-frequency analysis. Adaptive mode decomposition methods are the most apt strategy under these circumstances. Here, a shortlist of three well-established algorithms has been selected for an in-depth analysis. These signal decomposition approaches-namely, the Empirical Mode Decomposition (EMD), the Hilbert Vibration Decomposition (HVD), and the Variational Mode Decomposition (VMD)-are deemed to be the most representative ones because of their extensive use and favourable reception from the research community. The main aspects and properties of these data-adaptive methods, as well as their advantages, limitations, and drawbacks, are discussed and compared. Then, the potentialities of the three algorithms are assessed firstly on a numerical case study and then on a well-known experimental benchmark, including nonlinear cases and nonstationary signals.

Shape Sensing of Plate Structures Using the Inverse Finite Element Method: Investigation of Efficient Strain-Sensor Patterns.

Methods for real-time reconstruction of structural displacements using measured strain data is an area of active research due to its potential application for Structural Health Monitoring (SHM) and morphing structure control. The inverse Finite Element Method (iFEM) has been shown to be well suited for the full-field reconstruction of displacements, strains, and stresses of structures instrumented with discrete or continuous strain sensors. In practical applications, where the available number of sensors may be limited, the number and sensor positions constitute the key parameters. Understanding changes in the reconstruction quality with respect to sensor position is generally difficult and is the aim of the present work. This paper attempts to supplement the current iFEM modeling knowledge through a rigorous evaluation of several strain-sensor patterns for shape sensing of a rectangular plate. Line plots along various sections of the plate are used to assess the reconstruction quality near and far away from strain sensors, and the nodal displacements are studied as the sensor density increases. The numerical results clearly demonstrate the effectiveness of the strain sensors distributed along the plate boundary for reconstructing relatively simple displacement patterns, and highlight the potential of cross-diagonal strain-sensor patterns to improve the displacement reconstruction of more complex deformation patterns.

Phenotype characterisation of TBX4 mutation and deletion carriers with neonatal and paediatric pulmonary hypertension.

Rare variants in the T-box transcription factor 4 gene (TBX4) have recently been recognised as an emerging cause of paediatric pulmonary hypertension (PH). Their pathophysiology and contribution to persistent pulmonary hypertension in neonates (PPHN) are unknown. We sought to define the spectrum of clinical manifestations and histopathology associated with TBX4 variants in neonates and children with PH.We assessed clinical data and lung tissue in 19 children with PH, including PPHN, carrying TBX4 rare variants identified by next-generation sequencing and copy number variation arrays.Variants included six 17q23 deletions encompassing the entire TBX4 locus and neighbouring genes, and 12 likely damaging mutations. 10 infants presented with neonatal hypoxic respiratory failure and PPHN, and were subsequently discharged home. PH was diagnosed later in infancy or childhood. Three children died and two required lung transplantation. Associated anomalies included patent ductus arteriosus, septal defects, foot anomalies and developmental disability, the latter with a higher prevalence in deletion carriers. Histology in seven infants showed abnormal distal lung development and pulmonary hypertensive remodelling.TBX4 mutations and 17q23 deletions underlie a new form of developmental lung disease manifesting with severe, often biphasic PH at birth and/or later in infancy and childhood, often associated with skeletal anomalies, cardiac defects, neurodevelopmental disability and other anomalies.

Using Video Processing for the Full-Field Identification of Backbone Curves in Case of Large Vibrations.

Nonlinear modal analysis is a demanding yet imperative task to rigorously address real-life situations where the dynamics involved clearly exceed the limits of linear approximation. The specific case of geometric nonlinearities, where the effects induced by the second and higher-order terms in the strain-displacement relationship cannot be neglected, is of great significance for structural engineering in most of its fields of application-aerospace, civil construction, mechanical systems, and so on. However, this nonlinear behaviour is strongly affected by even small changes in stiffness or mass, e.g., by applying physically-attached sensors to the structure of interest. Indeed, the sensors placement introduces a certain amount of geometric hardening and mass variation, which becomes relevant for very flexible structures. The effects of mass loading, while highly recognised to be much larger in the nonlinear domain than in its linear counterpart, have seldom been explored experimentally. In this context, the aim of this paper is to perform a noncontact, full-field nonlinear investigation of the very light and very flexible XB-1 air wing prototype aluminum spar, applying the well-known resonance decay method. Video processing in general, and a high-speed, optical target tracking technique in particular, are proposed for this purpose; the methodology can be easily extended to any slender beam-like or plate-like element. Obtained results have been used to describe the first nonlinear normal mode of the spar in both unloaded and sensors-loaded conditions by means of their respective backbone curves. Noticeable changes were encountered between the two conditions when the structure undergoes large-amplitude flexural vibrations.

Trans-heterozygosity for mutations enhances the risk of recurrent/chronic pancreatitis in patients with Cystic Fibrosis.

BACKGROUND: Recurrent (RP) and chronic pancreatitis (CP) may complicate Cystic Fibrosis (CF). It is still unknown if mutations in genes involved in the intrapancreatic activation of trypsin (IPAT) or in the pancreatic secretion pathway (PSP) may enhance the risk for RP/CP in patients with CF. METHODS: We enrolled: 48 patients affected by CF complicated by RP/CP and, as controls 35 patients with CF without pancreatitis and 80 unrelated healthy subjects. We tested a panel of 8 genes involved in the IPAT, i.e. PRSS1, PRSS2, SPINK1, CTRC, CASR, CFTR, CTSB and KRT8 and 23 additional genes implicated in the PSP. RESULTS: We found 14/48 patients (29.2%) with mutations in genes involved in IPAT in the group of CF patients with RP/CP, while mutations in such genes were found in 2/35 (5.7%) patients with CF without pancreatitis and in 3/80 (3.8%) healthy subjects (p < 0.001). Thus, we found mutations in 12 genes of the PSP in 11/48 (22.9%) patients with CF and RP/CP. Overall, 19/48 (39.6%) patients with CF and RP/CP showed one or more mutations in the genes involved in the IPAT and in the PSP while such figure was 4/35 (11.4%) for patients with CF without pancreatitis and 11/80 (13.7%) for healthy controls (p < 0.001). CONCLUSIONS: The trans-heterozygous association between CFTR mutations in genes involved in the pathways of pancreatic enzyme activation and the pancreatic secretion may be risk factors for the development of recurrent or chronic pancreatitis in patients with CF.

Interstitial 10q21.1q23.31 Duplication due to Meiotic Recombination of a Paternal Balanced Complex Rearrangement: Cytogenetic and Molecular Characterization.

Complex chromosomal rearrangements (CCRs) are structural aberrations involving more than 2 chromosomal breakpoints. They are associated with different outcomes depending on the deletion/duplication of genomic material, gene disruption, or position effects. Balanced CCRs can also undergo missegregation during meiotic division, leading to unbalanced derivative chromosomes and, in some cases, to affected offspring. We report on a patient presenting with developmental and speech delay, growth retardation, microcephaly, hypospadias, and dysmorphic features, harboring an interstitial 10q21.1q23.31 duplication, due to recombination of a paternal CCR. Application of several cytogenetic and molecular techniques allowed determining the biological bases of the rearrangement, understanding the underlying chromosomal mechanism, and assessing the reproductive risk.

Extensive molecular analysis suggested the strong genetic heterogeneity of idiopathic chronic pancreatitis.

Rationale: Genetic features of Chronic Pancreatitis (CP) have been extensively investigated mainly testing genes associated to the trypsinogen activation pathway. However, different molecular pathways involving other genes may be implicated in CP pathogenesis. Objectives: 80 patients with Idiopathic CP were investigated using Next Generation Sequencing approach with a panel of 70 genes related to six different pancreatic pathways: premature activation of trypsinogen; modifier genes of Cystic Fibrosis phenotype; pancreatic secretion and ion homeostasis; Calcium signalling and zymogen granules exocytosis; autophagy; autoimmune pancreatitis related genes. Results: We detected mutations in 34 out of 70 genes examined; 64/80 patients (80.0%) were positive for mutations in one or more genes, 16/80 patients (20.0%) had no mutations. Mutations in CFTR were detected in 32/80 patients (40.0%) and 22 of them exhibited at least one mutation in genes of other pancreatic pathways. Of the remaining 48 patients, 13/80 (16.3%) had mutations in genes involved in premature activation of trypsinogen and 19/80 (23.8%) had mutations only in genes of the other pathways: 38/64 patients positive for mutations showed variants in two or more genes (59.3%). Conclusions: Our data, although to be extended with functional analysis of novel mutations, suggest a high rate of genetic heterogeneity in chronic pancreatitis and that trans-heterozygosity may predispose to the idiopathic CP phenotype.

Advantages of a next generation sequencing targeted approach for the molecular diagnosis of retinoblastoma.

BACKGROUND: Retinoblastoma (RB) is the most common malignant childhood tumor of the eye and results from inactivation of both alleles of the RB1 gene. Nowadays RB genetic diagnosis requires classical chromosome investigations, Multiplex Ligation-dependent Probe Amplification analysis (MLPA) and Sanger sequencing. Nevertheless, these techniques show some limitations. We report our experience on a cohort of RB patients using a combined approach of Next-Generation Sequencing (NGS) and RB1 custom array-Comparative Genomic Hybridization (aCGH). METHODS: A total of 65 patients with retinoblastoma were studied: 29 cases of bilateral RB and 36 cases of unilateral RB. All patients were previously tested with conventional cytogenetics and MLPA techniques. Fifty-three samples were then analysed using NGS. Eleven cases were analysed by RB1 custom aCGH. One last case was studied only by classic cytogenetics. Finally, it has been tested, in a lab sensitivity assay, the capability of NGS to detect artificial mosaicism series in previously recognized samples prepared at 3 different mosaicism frequencies: 10, 5, 1 %. RESULTS: Of the 29 cases of bilateral RB, 28 resulted positive (96.5 %) to the genetic investigation: 22 point mutations and 6 genomic rearrangements (four intragenic and two macrodeletion). A novel germline intragenic duplication, from exon18 to exon 23, was identified in a proband with bilateral RB. Of the 36 available cases of unilateral RB, 8 patients resulted positive (22 %) to the genetic investigation: 3 patients showed point mutations while 5 carried large deletion. Finally, we successfully validated, in a lab sensitivity assay, the capability of NGS to accurately measure level of artificial mosaicism down to 1 %. CONCLUSIONS: NGS and RB1-custom aCGH have demonstrated to be an effective combined approach in order to optimize the overall diagnostic procedures of RB. Custom aCGH is able to accurately detect genomic rearrangements allowing the characterization of their extension. NGS is extremely accurate in detecting single nucleotide variants, relatively simple to perform, cost savings and efficient and has confirmed a high sensitivity and accuracy in identifying low levels of artificial mosaicisms.

A De Novo CaSR Missense Variant in Combination with Two Inherited Missense Variants in CFTR and SPINK1 Detected in a Patient with Chronic Pancreatitis.

Chronic pancreatitis is often secondary to alcohol abuse, but pancreatitis with no other aetiology is frequently associated with variants in genes encoding proteins related to zymogen granule activation. Our goal was to identify genomic variants in a patient by analyzing an extended panel of genes associated with the intra-pancreatic activation of the trypsin pathway. A 23-year-old woman was addressed at our institution because of chronic pancreatitis of unknown aetiology presenting recurrent episodes since she was the age of four. Next Generation Sequencing was performed to analyze a panel of nine genes associated with pancreatitis (CaSR, CFTR, CPA1, CTRC, CTSB, KRT8, PRSS1, PRSS2, and SPINK1). Three missense variants were found: p.Leu997Phe, maternally inherited, in the CFTR gene; p.Ile73Phe, paternally inherited, in the SPINK1 gene; and p.Phe790Ser, a de novo variant, in the CaSR gene. They were classified, respectively as probably benign, a Variant of Uncertain Significance, and the last one, which has never been described in the literature, as likely being pathogenic following American College of Medical Genetics and Genomics standard guidelines. Extensive intra-pancreatic activation of trypsin pathway gene sequencing detected rare variants that were not found with other gene screening and showed that variants in different genes may interact in contributing to the onset of the pancreatitis phenotype.

Gene amplification as double minutes or homogeneously staining regions in solid tumors: origin and structure.

Double minutes (dmin) and homogeneously staining regions (hsr) are the cytogenetic hallmarks of genomic amplification in cancer. Different mechanisms have been proposed to explain their genesis. Recently, our group showed that the MYC-containing dmin in leukemia cases arise by excision and amplification (episome model). In the present paper we investigated 10 cell lines from solid tumors showing MYCN amplification as dmin or hsr. Particularly revealing results were provided by the two subclones of the neuroblastoma cell line STA-NB-10, one showing dmin-only and the second hsr-only amplification. Both subclones showed a deletion, at 2p24.3, whose extension matched the amplicon extension. Additionally, the amplicon structure of the dmin and hsr forms was identical. This strongly argues that the episome model, already demonstrated in leukemias, applies to solid tumors as well, and that dmin and hsr are two faces of the same coin. The organization of the duplicated segments varied from very simple (no apparent changes from the normal sequence) to very complex. MYCN was always overexpressed (significantly overexpressed in three cases). The fusion junctions, always mediated by nonhomologous end joining, occasionally juxtaposed truncated genes in the same transcriptional orientation. Fusion transcripts involving NBAS (also known as NAG), FAM49A, BC035112 (also known as NCRNA00276), and SMC6 genes were indeed detected, although their role in the context of the tumor is not clear.

Design and Mechanical Characterization Using Digital Image Correlation of Soft Tissue-Mimicking Polymers.

Present and future anatomical models for biomedical applications will need bio-mimicking three-dimensional (3D)-printed tissues. These would enable, for example, the evaluation of the quality-performance of novel devices at an intermediate step between ex-vivo and in-vivo trials. Nowadays, PolyJet technology produces anatomical models with varying levels of realism and fidelity to replicate organic tissues. These include anatomical presets set with combinations of multiple materials, transitions, and colors that vary in hardness, flexibility, and density. This study aims to mechanically characterize multi-material specimens designed and fabricated to mimic various bio-inspired hierarchical structures targeted to mimic tendons and ligaments. A Stratasys® J750™ 3D Printer was used, combining the Agilus30™ material at different hardness levels in the bio-mimicking configurations. Then, the mechanical properties of these different options were tested to evaluate their behavior under uni-axial tensile tests. Digital Image Correlation (DIC) was used to accurately quantify the specimens' large strains in a non-contact fashion. A difference in the mechanical properties according to pattern type, proposed hardness combinations, and matrix-to-fiber ratio were evidenced. The specimens V, J1, A1, and C were selected as the best for every type of pattern. Specimens V were chosen as the leading combination since they exhibited the best balance of mechanical properties with the higher values of Modulus of elasticity (2.21 ± 0.17 MPa), maximum strain (1.86 ± 0.05 mm/mm), and tensile strength at break (2.11 ± 0.13 MPa). The approach demonstrates the versatility of PolyJet technology that enables core materials to be tailored based on specific needs. These findings will allow the development of more accurate and realistic computational and 3D printed soft tissue anatomical solutions mimicking something much closer to real tissues.

Implantable medical devices for tendon and ligament repair: a review of patents and commercial products.

INTRODUCTION: Tendon and ligament injuries are a frequent and debilitating issue that affects many patients worldwide. The predominant solution is the suture thread, which is not without potential side effects and limitations. Implantable medical devices have gained more attention as an alternative approach. However, due to the many challenges of the inner body environment (limited available space, chemically aggressive environment, etc), the development of suitable devices is not exempt from practical and technical difficulties. AREAS COVERED: Here, implantable medical devices for tendon and ligaments injuries are reviewed and discussed. Commercially-available products and registered patents are all considered as long as they fit the standard definitions of 'implantable medical devices' (reported in the Introduction). The research was then narrowed down to five commercial products, deemed as the most representative of the whole market. Their effectiveness and performance are analysed, as well as the possible areas of improvement and development. EXPERT OPINION: Commercially available products present overall superior mechanical performances than suture techniques. Nevertheless, these latter ones might be still preferred for their wider range of customization. This aspect, and many others, could represent an area of improvement for implantable medical devices, to further explore their potential for tendon and ligament repair.

3D printing and testing of rose thorns or limpet teeth inspired anchor device for tendon tissue repair.

PURPOSES: Advancements in medical technology have enabled medical specialists to resolve significant problems concerning tendon injuries. However, despite the latest improvements, surgical tendon repair remains challenging. This study aimed to explore the capabilities of the current state-of-the-art technologies for implantable devices. METHODS: After performing extensive patent landscaping and literature review, an anchored tissue fixation device was deemed the most suitable candidate. This design was firstly investigated numerically, realizing a Finite Element Model of the device anchored to two swine tendons stumps, to simulate its application on a severed tendon. Two different hook designs, both bio-inspired, were tested while retaining the same device geometry and anchoring strategy. Then, the applicability of a 3D-printed prototype was tested on swine tendons. Finally, the device-tendon stumps ensemble was subjected to uniaxial tensile tests. RESULTS: The results show that the investigated device enables a better load distribution during the immobilized limb period in comparison to standard suture-based approaches, yet it still presents several design flaws. CONCLUSIONS: The current implantable solutions do not ensure an optimal result in terms of strength recovery. This and other weak points of the currently available proposals will serve as a starting point for future works on bio-inspired implantable devices for tendon repair.

Effects of the Manufacturing Methods on the Mechanical Properties of a Medical-Grade Copolymer Poly(L-lactide-co-D,L-lactide) and Poly(L-lactide-co-ε-caprolactone) Blend.

Biocompatible and biodegradable polymers represent the future in the manufacturing of medical implantable solutions. As of today, these are generally manufactured with metallic components which cannot be naturally absorbed within the human body. This requires performing an additional surgical procedure to remove the remnants after complete rehabilitation or to leave the devices in situ indefinitely. Nevertheless, the biomaterials used for this purpose must satisfy well-defined mechanical requirements. These are difficult to ascertain at the design phase since they depend not only on their physicochemical properties but also on the specific manufacturing methods used for the target application. Therefore, this research was focused on establishing the effects of the manufacturing methods on both the mechanical properties and the thermal behavior of a medical-grade copolymer blend. Specifically, Injection and Compression Molding were considered. A Poly(L-lactide-co-D,L-lactide)/Poly(L-lactide-co-ε-caprolactone) blend was considered for this investigation, with a ratio of 50/50 (w/w), aimed at the manufacturing of implantable devices for tendon repair. Interesting results were obtained.

Clinical hallmarks and genetic polymorphisms in the CFTR gene contribute to the disclosure of the A1006E mutation.

Since the identification of the Cystic Fibrosis transmembrane conductance regulator (CFTR) gene in 1989, many genetic mutations have been found in cystic fibrosis (CF) patients. Dysfunctions of the CFTR gene are responsible for the highly variable clinical presentation ranging from severe CF, disseminated bronchiectasis, idiopathic chronic pancreatitis and congenital bilateral absence of vas deferens (CBAVD). Linkage disequilibrium studies have shown that some mutations are stringently coupled with polymorphisms in a genetic complex called haplotype. From a familial study of a patient with CBAVD, carrier of the A1006E mutation, we have observed its strict association with the polymorphism 5T-TG11. In order to speed up the genetic diagnosis and to correlate the clinical setting to this genetic feature, we have directly investigated the exon 17a, where the A1006E mutation is located, of five cystic fibrosis patients belonging to two unrelated families. All patients had the 5T-TG11 tract, F508del and one unknown mutation. One more family with two affected individuals carrying the Q220X/A1006E mutations was investigated for the poly-T polymorphism. All the members were found to have the A1006E mutation and the 5T-TG11 in the same DNA strand, demonstrating that this strategy is a reliable and inexpensive method for genotyping the CFTR gene. A detailed description of the clinical presentation and follow-up are provided in order to highlight common phenotypic features useful to improve the management of cystic fibrosis patients.

Lyonization effects of the t(X;16) translocation on the phenotypic expression in a rare female with Menkes disease.

Menkes disease (MD) is a rare and severe X-linked recessive disorder of copper metabolism. The MD gene, ATP7A (ATPase Cu++ transporting alpha polypeptide), encodes an ATP-dependent copper-binding membrane protein. In this report, we describe a girl with typical clinical features of MD, carrying a balanced translocation between the chromosomes X and 16 producing the disruption of one copy of ATP7A gene and the silencing of the other copy because of the chromosome X inactivation. Fluorescence in situ hybridization experiments with bacterial derived artificial chromosome probes revealed that the breakpoints were located within Xq13.3 and 16p11.2. Replication pattern analysis demonstrated that the normal X chromosome was late replicating and consequently inactivated, whereas the der(X)t(X;16), bearing the disrupted ATP7A gene, was active. An innovative approach, based on FMR1 (fragile X mental retardation 1) gene polymorphism, has been used to disclose the paternal origin of the rearrangement providing a new diagnostic tool for determining the parental origin of defects involving the X chromosome and clarifying the mechanism leading to the cytogenetic rearrangement that occurred in our patient.

Novel Compound Heterozygous Mutations in IL-7 Receptor α Gene in a 15-Month-Old Girl Presenting With Thrombocytopenia, Normal T Cell Count and Maternal Engraftment.

Patients with severe combined immunodeficiency (SCID) exhibit T lymphopenia and profound impairments in cellular and humoral immunity. IL-7 receptor α (IL-7Rα) deficiency is a rare form of SCID that usually presents in the first months of life with severe and opportunistic infections, failure to thrive and high risk of mortality unless treated. Here, we reported an atypical and delayed onset of IL7Rα-SCID in a 15-month-old girl presenting with thrombocytopenia. Immunological investigations showed a normal lymphocyte count with isolated CD4-penia, absence of naïve T cells, marked hypergammaglobulinemia, and maternal T cell engraftment. Targeted next generation sequencing (NGS) revealed two novel compound heterozygous mutations in the IL-7Rα gene: c.160T>C (p.S54P) and c.245G>T (p.C82F). The atypical onset and the unusual immunological phenotype expressed by our patient highlights the diagnostic challenge in the field of primary immunodeficiencies (PID) and in particular in SCID patients where prompt diagnosis and therapy greatly affects survival.