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Background & Aims: About 15% of intrahepatic cholangiocarcinomas (iCCAs) express fibroblast growth factor receptor 2 (FGFR2) fusion proteins (FFs), usually alongside mutational inactivation of TP53, CDKN2A or BAP1. In FFs, FGFR2 residues 1-768 fuse to sequences encoded by a diverse array of partner genes (>60) causing oncogenic FF activation. While FGFR-specific tyrosine kinase inhibitors (F-TKI) provide clinical benefit in FF + iCCA, responses are partial and/or limited by resistance mechanisms, such as the V565F substitution in the FGFR2 gatekeeper residue. Improving on FF targeting in iCCA therefore remains a critical unmet need. Herein, we aimed to generate a murine model of FF-driven iCCA and use this to uncover actionable FF-associated dependencies., Methods: Four iCCA FFs carrying different fusion sequences were expressed in Tp53 -/- mouse liver organoids. Tumorigenic properties of genetically modified liver organoids were assessed by transplantation into immuno-deficient mice. Cellular models derived from neoplastic lesions were exploited for pre-clinical studies., Results: Transplantation of FF-expressing liver organoids yielded tumors diagnosed as CCA based on histological, phenotypic and transcriptomic analyses. The penetrance of this tumorigenic phenotype was influenced by FF identity. Tumor organoids and 2D cell lines derived from CCA lesions were addicted to FF signaling via Ras-Erk, regardless of FF identity or V565F mutation. Dual blockade of FF and the Ras-Erk pathway by concomitant pharmacological inhibition of FFs and Mek1/2 provided greater therapeutic efficacy than single agent F-TKI in vitro and in vivo., Conclusions: FF-driven iCCA pathogenesis was successfully modeled on a Tp53 -/- murine background, revealing biological heterogeneity among structurally different FFs. Double blockade of FF-ERK signaling deserves consideration for precision-based approaches against human FF + iCCA., Lay Summary: Intrahepatic cholangiocarcinoma (iCCA) is a rare cancer that is difficult to treat. A subtype of iCCA is caused by genomic alterations that generate oncogenic drivers known as FGFR2 fusions. Patients with FGFR2 fusions respond to FGFR inhibitors, but clinical responses are often of modest duration. We used animal and cellular models to show that FGFR2 fusions require the activity of a downstream effector named Mek1/2. We found that dual blockade of FGFR2 fusions and Mek1/2 was more effective than isolated inhibition of FGFR2 fusions, pointing to the potential clinical utility of dual FGFR2-MEK1/2 blockade in patients with iCCA., Competing Interests: Conflict of interest All Authors, except M.J.B., have no personal, professional or financial conflicts to disclose. M.J.B. disclosures: ADC Therapeutics – Consulting to self; Exelixis Pharmaceuticals – Consulting to self; Inspyr Therapeutics – Consulting to self; G1 Therapeutics – Consulting to self; Immunovative Therapies – Consulting to self; OncBioMune Pharmaceuticals – Consulting to self; Western Oncolytics – Consulting to self; Lynx Group – Consulting to self; Genentech – Consulting to self; Merck – Consulting to self; Huya – Consulting to self; Astra Zeneca – Travel Support to self. Please refer to the accompanying ICMJE disclosure forms for further details., (Copyright © 2021 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

The first wave of COVID-19 spread worldwide from March to May 2020. Italy was one of the countries in the world where the lockdown period was most prolonged and restrictive. To date, the impact of prolonged lockdown on pediatric traumas has not fully investigated. This paper aimed to analyze, and compare to 2019, the incidence and the fracture pattern in patients admitted to our pediatric hospital during the total lockdown period. A single-center retrospective study was performed. The data were gathered from the Pediatric Emergency Department (PED) of the Bambino Gesù Children Hospital of Rome (Palidoro). This PED is the pediatric referral center for Rome and the hub for pediatric traumas of the region. Any admission diagnosis for fracture, trauma, sprains and dislocation during the lockdown period (10 March-4 May) were included. The demographic data, diagnosis, type of treatment, fracture segment, bone involvement and time interval between trauma and presentation to the PED were recorded. In 2020, a reduction of total traumas and fractures, compared to 2019 ( p < 0.001), occurred (81%). Superior limb and inferior limb fractures decreased in 2020 compared to 2019 ( p < 0.05). The identification of pediatric traumas and fractures trend could be useful to reorganize the PED. Epidemiological data from the previous lockdown could be helpful to prepare the healthcare system for new pandemic waves. Moreover, sharing national statistics and correlating those to other countries' protocols, could be helpful to solve problems in case of worldwide emergency situations.

Variants in tank-binding kinase 1 (TBK1) are responsible for a significant proportion of amyotrophic lateral sclerosis (ALS) cases. In the present study, we analyzed variants in TBK1 extracted by targeted sequencing of 32 genes in a group of 406 Italian patients with ALS. We identified 7 different TBK1 variants in 7 sporadic cases, resulting in a frequency of 1.7%. Three patients had missense variants (p.R357Q, p.R358H, and p.R724C), one patient had a small deletion (p.E618del), and 3 had truncating variants (p.Y482*, p.R229*, and p.N681*). Notably, we found that 4 patients had an additional variant in ALS-related genes: 2 in OPTN and 2 in the 3'UTR region of FUS. By studying an independent group of 7 TBK1-mutated patients previously reported, we found another variant in the 3'UTR region of FUS in one patient. The presence of a second variant in TBK1 variant carriers is an interesting finding that needs to be investigated in larger cohorts of patients. These findings suggest that TBK1 belongs to the category of genes conferring a significantly increased risk but not sufficient to cause disease., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The aim of this study is to implement the clinical use of the three-dimensional (3D) design and printing technology in pediatric pathologies requiring immobilization. We describe the manufacturing process of the 3D device in place of the plaster cast usually applied to a child 48/72 h after the access to the Trauma Center Traumatology Hub. This procedure had already been performed at Level II, Trauma Center, Campania Region, Orthopaedic Division of Santobono Children's Hospital, Naples, Italy. The operative phase was performed by two 3D printers and a scanner in the bioengineering laboratory of the hospital's outpatient area. The phase of software elaboration requires close cooperation among physicians and engineers. We decided to use a model with a double-shell design and holes varying in width to ensure complete ventilation and lightness of the device. We chose to treat nondisplaced metaphyseal distal fractures of the radius in 18 patients enrolled from January 2017 to November 2017. The flow chart includes clinical and radiological examinations of every enrolled child, collecting information required by the program and its elaboration by bioengineers, and then transfer of the results to 3D printers. The child, immobilized by a temporary splint, wore his 3D device after 12/24 h. Then, he underwent serial check-ups in which the effectiveness and appropriateness of the treatment were clinically monitored and evaluated using subjective scales: visual analogue scale and patient-rated wrist evaluation. All the fractures consolidated both radiologically and clinically after the treatment, with no complications reported. Only one partial breakage of the device happened because of an accidental fall. The statistical analysis of the visual analogue scale and patient-rated wrist evaluation data shows that children's activities of everyday life improved during the immobilization thanks to this treatment. This first study shows that using a 3D device instead of a traditional plaster cast can be an effective alternative approach in the treatment of pediatric nondisplaced metaphyseal distal radius fractures, with high overall patient satisfaction. We believe that 3D technology could be extended to the treatment of more complex fractures; this will be the subject of our second study.

About 15% of intrahepatic cholangiocarcinomas (ICCs) express constitutively active fibroblast growth factor receptor 2 (FGFR2) fusion proteins (FFs) generated by chromosomal translocations. FFs have been nominated as oncogenic drivers because administration of FGFR tyrosine kinase inhibitors (F-TKIs) can elicit meaningful objective clinical responses in patients carrying FF-positive ICC. Thus, optimization of FF targeting is a pressing clinical need. Herein, we report that three different FFs, previously isolated from ICC samples, are heat shock protein 90 (HSP90) clients and undergo rapid degradation upon HSP90 pharmacological blockade by the clinically advanced HSP90 inhibitor ganetespib. Combining catalytic suppression by the F-TKI BGJ398 with HSP90 blockade by ganetespib suppressed FGFR2-TACC3 (transforming acidic coiled-coil containing protein 3) signaling in cultured cells more effectively than either BGJ398 or ganetespib in isolation. The BGJ398 + ganetespib combo was also superior to single agents when tested in mice carrying subcutaneous tumors generated by transplantation of FGFR2-TACC3 NIH3T3 transformants. Of note, FF mutants known to enforce clinical resistance to BGJ398 in ICC patients retained full sensitivity to ganetespib in cultured cells. Conclusion: Our data provide a proof of principle that upfront treatment with the BGJ398 + ganetespib combo improves therapeutic targeting of FGFR2 fusions in an experimental setting, which may be relevant to precision medicine approaches to FF-driven ICC., (© 2018 by the American Association for the Study of Liver Diseases.)

Cutaneous ulceration is a difficult medical problem and a major source of morbidity for patients. In the surgical treatment of ulcers, debridement is the first step, and it can be carried out using several surgical tools. Recently, new surgical devices have emerged using plasma-mediated electrical discharges with a lower peak temperature. A prospective single-blind trial was conducted on chronic ulcers not responsive to common non-surgical management. Patients were randomly separated into 2 groups: Group A received surgical debridement with conventional electrocautery, and Group B received surgical debridement using the plasma-mediated device. Histological samples were collected intraoperatively to evaluate the thermal damage during the surgical procedure and 2 weeks after surgery to evaluate the inflammatory response and collagen deposition. The width of coagulation necrosis at the incision margins in Group B was significantly shorter compared with Group A (P = .001). The inflammatory cell infiltration showed a cellular distribution percentage that was quite equal between the 2 groups. The granulation tissue showed an abundant deposition of dense and mature collagen in Group B, compared with Group A, where the mature collagen appeared in small quantities (P < .001). Microbial culture showed a lower incidence of postoperative infections in Group B compared with the control group (P < .05). The study demonstrated, based on the results, that the new technology with the use of a lower temperature electrosurgical device represents an effective therapeutic weapon for the surgical treatment of skin ulcers, both vascular and extravascular types., (© 2018 Medicalhelplines.com Inc and John Wiley & Sons Ltd.)

The use of Total Knee Arthroplasty (TKA) in treatment of chronic degenerative pathologies of the knee boasts of an experience of 50 years. During this period the collaboration between surgeons and engineers produced many developments in the design of the prosthesis. Today this procedure is safe and established even if in continuous development. The progress in technologies and the use of new materials let researches try again old-fashioned techniques from the past in order to be improved. This enthusiasm for those discovers is not always going hand to hand with scientific validation: many open questions remains Every different concept of the design tries to answer to special needs as the reach of the highest ROM, the reduction of pain and debris, articular geometry, the type of fixation, the modularity of augments and stems, the types of constraints,  knee kinematic and of course costs.

Background: Venous thromboembolism, a spectrum of diseases ranging from deep venous thrombosis to pulmonary embolism, is a major source of morbidity and mortality. The majority of cases described in plastic surgery involve abdominoplasty. Risk assessment and prophylaxis are paramount in such patients. General recommendations were recently developed, but the evidence in the literature was insufficient to prepare exhaustive guidelines regarding the medication, dosage, timing, or length of the prophylaxis., Methods: A thromboprophylaxis protocol was developed for patients undergoing abdominoplasty. The protocol consisted of preoperative, intraoperative, and postoperative measures. Enoxaparin was administered as chemoprophylaxis in selected patients. The study involved 253 patients. The patients were analyzed for age, body mass index, enoxaparin dosage, risk factors, and complications., Results: Deep venous thrombosis was documented in two cases (0.8%). No pulmonary embolism occurred. Three patients (1.2%) presented mild subcutaneous abdominal hematoma within the first postoperative week that spontaneously resorbed with neither aesthetic nor functional complications. Two patients (0.8%) presented severe hematoma requiring surgical re-intervention for drainage and hemostasis revision. Statistical analysis showed no significant correlation between enoxaparin dosage and hematoma (P=0.18) or deep venous thrombosis (P=0.61)., Conclusions: The described thromboprophylaxis protocol proved to be effective in the prevention of thrombotic events, with an acceptable risk of hemorrhagic complications. Furthermore, it provides new evidence regarding the currently debated variables of chemoprophylaxis, namely type, dosage, timing, and length.

The ErbB signaling network instructs the execution of key cellular programs, such as cell survival, proliferation and motility, through the generation of robust signals of defined strength and duration. In contrast, unabated ErbB signaling disrupts tissue homeostasis and leads to cell transformation. Cells oppose the threat inherent in excessive ErbB activity through several mechanisms of negative feedback regulation. Inducible feedback inhibitors (IFIs) are expressed in the context of transcriptional responses triggered by ErbB signaling, thus being uniquely suited to regulate ErbB activity during the execution of complex cellular programs. This review focuses on MIG6, an IFI that restrains ErbB signaling by mediating ErbB kinase suppression and receptor down-regulation. We will review key issues in MIG6 function, regulation and tumor suppressor activity. Subsequently, the role for MIG6 loss in the pathogenesis of tumors driven by ErbB oncogenes as well as in the generation of cellular addiction to ErbB signaling will be discussed. We will conclude by analyzing feedback inhibition by MIG6 in the context of therapies directed against ErbB and non-ErbB oncogenes., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

GDNF is a Sertoli-cell-derived factor that controls the balance between self-renewal and differentiation of the spermatogonial stem cells. Although research in recent years has concentrated on the impact of GDNF on target germ cells rather little attention has been paid to the molecular control of GDNF expression in Sertoli cells. Here, we aimed to characterize the promoter region of the mouse gdnf gene active in Sertoli cells. We identified the transcriptional start sites and analyzed the promoter activity of the 5'-flanking regions. By in-silico analysis of evolutionarily conserved DNA sequences we identified several putative transcription factor-binding regions. Deletion analysis showed the involvement of the three CRE sites for basal and cAMP-induced expression of gdnf in murine Sertoli cells. These results provide the basis for future studies to analyze how hormonal or paracrine signals modulate the transcriptional activity of gdnf in Sertoli cells., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

In mammals, the biological activity of the stem/progenitor compartment sustains production of mature gametes through spermatogenesis. Spermatogonial stem cells and their progeny belong to the class of undifferentiated spermatogonia, a germ cell population found on the basal membrane of the seminiferous tubules. A large body of evidence has demonstrated that glial cell line-derived neurotrophic factor (GDNF), a Sertoli-derived factor, is essential for in vivo and in vitro stem cell self-renewal. However, the mechanisms underlying this activity are not completely understood. In this study, we show that GDNF induces dose-dependent directional migration of freshly selected undifferentiated spermatogonia, as well as germline stem cells in culture, using a Boyden chamber assay. GDNF-induced migration is dependent on the expression of the GDNF co-receptor GFRA1, as shown by migration assays performed on parental and GFRA1-transduced GC-1 spermatogonial cell lines. We found that the actin regulatory protein vasodilator-stimulated phosphoprotein (VASP) is specifically expressed in undifferentiated spermatogonia. VASP belongs to the ENA/VASP family of proteins implicated in actin-dependent processes, such as fibroblast migration, axon guidance, and cell adhesion. In intact seminiferous tubules and germline stem cell cultures, GDNF treatment up-regulates VASP in a dose-dependent fashion. These data identify a novel role for the niche-derived factor GDNF, and they suggest that GDNF may impinge on the stem/progenitor compartment, affecting the actin cytoskeleton and cell migration.