A Solve-RD ClinVar-based reanalysis of 1522 index cases from ERN-ITHACA reveals common pitfalls and misinterpretations in exome sequencing

Purpose Within the Solve-RD project (https://solve-rd.eu/), the European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies aimed to investigate whether a reanalysis of exomes from unsolved cases based on ClinVar annotations could establish additional diagnoses. We present the results of the “ClinVar low-hanging fruit” reanalysis, reasons for the failure of previous analyses, and lessons learned. Methods Data from the first 3576 exomes (1522 probands and 2054 relatives) collected from European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies was reanalyzed by the Solve-RD consortium by evaluating for the presence of single-nucleotide variant, and small insertions and deletions already reported as (likely) pathogenic in ClinVar. Variants were filtered according to frequency, genotype, and mode of inheritance and reinterpreted. Results We identified causal variants in 59 cases (3.9%), 50 of them also raised by other approaches and 9 leading to new diagnoses, highlighting interpretation challenges: variants in genes not known to be involved in human disease at the time of the first analysis, misleading genotypes, or variants undetected by local pipelines (variants in off-target regions, low quality filters, low allelic balance, or high frequency). Conclusion The “ClinVar low-hanging fruit” analysis represents an effective, fast, and easy approach to recover causal variants from exome sequencing data, herewith contributing to the reduction of the diagnostic deadlock

Orthopaedic concerns in children with growth hormone therapy.

Growth hormone (GH) therapy is widely used in children; it may have various severe orthopaedic complications. Slipped capital femoral epiphysis, Legg-Calvé-Perthes disease, scoliosis and carpal tunnel syndrome may occur with GH treatment. Before beginning GH therapy, it is important to take into account all the risk factors of the individual patient, as some conditions could contraindicate GH treatment. During GH treatment, close monitoring with both clinical and radiographic examination is mandatory. The paediatric orthopaedic surgeon will frequently be asked about the management of these complications and about the necessity for treatment arrest. The authors review the orthopaedic complications which the orthopaedic surgeon may encounter in patients treated with GH

Fetal umbilical-systemic shunt with a positive issue.

We herein report the case of abnormal umbilical-venous return in which the antenatal ultrasound enabled us to establish the diagnosis of umbilical-systemic shunt (Type 1 according to Achiron (Achiron and Kivilevitch, 2016)). Due to the concomitant associations of cardiomegaly, intrauterine growth retardation, oligohydramnios, and left-lobe hypoplasia with agenesis of the intrahepatic umbilical vein - left portal vein - ductus venosus, a poor prognosis (11.1% survival) was to be expected. In spite of development of pulmonary arterial hypertension at birth, which was promptly treated, the evolution was nevertheless good, both on clinical and ultrasound follow-up

Self-expanding metallic stents in benign postoperative biliary strictures: a difficult surgical obstacle?

BACKGROUND/AIMS: To report the hazards of surgical management in patients suffering from benign postoperative biliary strictures (BBS) previously treated with self-expanding metallic stents (SEMS). METHODOLOGY: Five patients with BBS were treated by SEMS (Wallstent in 3 patients and Gianturco stent in 2) through an endoscopic (2 patients) or a percutaneous transhepatic route (3 patients). Metallic stent obstruction due to intraprosthetic epithelial hyperplasia was observed in all patients, being responsible for severe cholangitis and intrahepatic stones in 3 and 2 patients respectively. RESULTS: One patient remains free of symptoms but presented with right posterior liver atrophy. Difficult biliary repair was encountered in 3 patients, requiring excision and reconstruction of the main biliary convergence for severely damaged biliary wall and epithelium by metallic stents. One patient with a SEMS extended upward the right secondary biliary divisions required a right hemihepatectomy. CONCLUSIONS: The surgical management of benign biliary strictures is severely complicated by the presence of metallic stents. Thus, SEMS should not be employed for treating BBS in healthy patients with a low operative risk

Suspicious myometrial mass on ultrasonography and MRI does not necessarily mean a sarcoma on histology

We report an unusual ultrasonographic and magnetic resonance imaging (MRI) presentation of a myometrial mass in a 38-year-old woman hoping to conceive. Hysterectomy had been proposed elsewhere because of the suspicious nature of the mass, but the patient was seeking a second opinion. This atypical formation looked consistent with either hydropic degeneration of a uterine myoma or leiomyosarcoma, but preoperative differential diagnosis was impossible. Laparoscopic tumorectomy was performed and histology confirmed a degenerating uterine myoma. We, therefore, show that unusual ultrasonographic and MRI findings do not necessarily require radical surgery, even if sarcoma cannot be excluded preoperatively, especially in patients who wish to conceiv

Quantification of endometriotic lesions in a murine model by fluorimetric and morphometric analyses.

BACKGROUND: In animal models of endometriosis, the identification and quantification of lesions originating from human endometrium is often hampered by the small size of the implants and their embedding in murine tissue. The purpose of the present study was to develop two new methods of quantifying endometriosis-like lesions in a nude mouse model: fluorimetry and morphometry. METHODS: Human menstrual endometrium was labelled using a fluorescent tracker, carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE), and transplanted into the pelvic cavity of mice by injection through the peritoneum after performing a cutaneous incision. After 5 days, lesions were recovered by laparotomy. The fluorescence of the recovered endometriotic lesions was measured. Endometrial stroma and glands were immunostained in lesion sections with anti-CD10 and anti-CK22 antibodies, and their surface area was evaluated by morphometric analysis. RESULTS: Fluorescent labelling allows identification of lesions not visible macroscopically. A good correlation was observed between fluorimetry and morphometry (r=0.88) applied for lesion quantification. CONCLUSIONS: Fluorimetric evaluation combined with morphometric analysis of endometriosis-like lesions allows objective and reliable recording of endometriosis development in a nude mouse model. This quantification method could therefore be useful for future pharmacological and toxicological studies