Genome-wide meta-analysis implicates variation affecting mast cell biology in urticaria.

BACKGROUND: Urticaria is characterized by inappropriate mast cell degranulation leading to the development of wheals and/or angioedema. Twin and family studies indicate that there is a substantial heritable component to urticaria risk. OBJECTIVE: Our aim was to identify genomic loci at which common genetic variation influences urticaria susceptibility. METHODS: Genome-wide association studies of urticaria (including all subtypes) from 3 European cohorts (UK Biobank, FinnGen, and the Trøndelag Health Study [HUNT]) were combined through statistical meta-analysis (14,306 urticaria cases and 650,664 controls). Cases were identified via electronic health care records from primary and/or secondary care. To identify putative causal variants and genes, statistical fine-mapping, colocalization, and interrogation of publicly available single-cell transcriptome sequencing resources were performed. RESULTS: Genome-wide significant associations (P < 5 × 10-8) were identified at 6 independent loci. These included 2 previously reported association signals at 1q44 and the human leucocyte antigen region on chromosome 6. Genes with expected or established roles in mast cell biology were associated with the 4 other genome-wide association signals (GCSAML, FCER1A, TPSAB1, and CBLB). Colocalization of association signals consistent with the presence of shared causal variants was observed between urticaria susceptibility and increased expression of GCSAML (posterior probability of colocalization [PPcoloc] = 0.89) and FCER1A (PPcoloc = 0.91) in skin. CONCLUSION: Common genetic variation influencing the risk of developing urticaria was identified at 6 genomic loci. The relationship between genes with roles in mast cell biology and several association signals implicates genetic variability of specific components of mast cell function in the development of urticaria.

New Trends in Neurosurgery: Toward a Future of Integration and Equity Among Male and Female Neurosurgeons.

The gradual rise of women in medical schools and residencies, surpassing men in medical school applications, contrasts with the male dominance in surgical fields, including neurosurgery (only 18% women). Reasons include concerns about work-life balance, traditional childcare roles, and gender biases. In response, Women in Neurosurgery (WINs) was founded in 1989 to address gender disparities. However, WINs sessions at conferences evolved into segregated scientific sessions, deviating from their original purpose. This contradicts the ideal of a unified neurosurgical community. While some support segregated spaces, many advocate against gender-based divisions. Today WINs' existence is deemed outdated, with a call for integration, inclusivity, and equality in the modern era of neurosurgery.

Computerized Three-Dimensional Analysis: A Novel Method to Assess the Effect of Open-Door Laminoplasty.

INTRODUCTION: The three-dimensional elaboration of morphological data derived from computed tomography (CT) and magnetic resonance imaging (MRI) scans generates virtual anatomical reconstructions. Here, we propose a novel protocol to analyze the postoperative results of open-door laminoplasty to evaluate differences in the volume of the spinal canal. The protocol uses geometric models in patients with cervical degenerative myelopathy before versus after cervical laminoplasty. MATERIALS AND METHODS: Mimics and 3-Matic software (Materialise, Leuven, BE) programs were used to segment anatomical structures and create polygon meshes of spines. Patients with cervical spondylotic myelopathy were enrolled. The models obtained before and after laminoplasty were superimposed by using a global registration function. The magnitude of divergence was quantified by using the root-mean-square error (RMSE). RESULTS: Using this novel protocol, we were able to map the differences in the volume of the spinal canal before laminoplasty and after laminoplasty and to quantify its magnitude and calculate the volumes. DISCUSSION AND CONCLUSIONS: The development of a procedure to measure the space within the cervical bone walls using geometric parameters represents a new, powerful method to verify the results obtained by cervical laminoplasty. Further research horizons may include the routine use of virtual models in surgical planning for this procedure.

Limits and usefulness of intraoperative evoked potentials during laminoplasty.

BACKGROUND: Over the past 10 years, intraoperative neurophysiological monitoring (IONM) has been widely performed during surgery for treating spondylotic cervical myelopathy. Our study considers the predictive value of IONM during laminoplasty, regarding, first, the adequacy of spinal cord decompression and, second, the long-term neuro-functional outcome. METHODS: We considered 38 patients with the diagnosis of degenerative cervical myelopathy who underwent an open-door laminoplasty. All patients were evaluated preoperatively, and at three and 12 months postoperatively, with the Japanese Orthopedic Association (JOA) point scale. Upper and lower limb somatosensory and motor evoked potentials (SSEPs and MEPs) were recorded preoperatively and intraoperatively. RESULTS: During surgery, three of 38 patients showed a deterioration of SSEPs and MEPs compared to baseline values. Surgery was then converted from laminoplasty to laminectomy, resulting in the gradual restoration of the evoked potentials. The neurophysiological parameter significantly associated with a better clinical outcome was the latency of lower limbs MEPs. The 12 patients who had a more prominent reduction of the MEPs latency at the end of surgery showed a higher post-surgical JOA score, increasing ≥30% compared to baseline values at the 3- and 12-month follow-up. CONCLUSIONS: Though not a predictor of clinical outcome, the IONM was essential to evaluate the effectiveness of spinal cord decompression. Reduced latency of lower limbs MEPs may predict a better clinical outcome. We suggest that IONM in patients with degenerative cervical myelopathy should be routine. It is necessary to conduct larger studies to clarify the predictive value of IONM.

Telemedicine in practice: Patient selection and strategies adopted in spinal care during the COVID-19 era and beyond.

Background: The SARS-CoV-2 (COVID-19) pandemic has had a substantial effect on health-care systems around the world. To deal with this challenge, we developed a prospectus design and test a teleconsultation procedure suitable for both diagnostic and therapeutic needs of outpatients in our Spine and Spinal Cord Surgery Units. Methods: (1) The first 15 patients received in-person and telemedicine visits: Our pilot began with screening protocol that involved 15 patients who received an in-person assessment and a telemedicine consult. The 15 patients who were selected were already known to our unit after having had at least one previous in-person visit. Further, they had to be neurologically intact or have a stable neurological examination. The secondary teleconsultation took place as a synchronous face-to-face communication between the doctor and the patient through a video interface (Lifesize Video Conferencing, Austin, Texas). If the patient demonstrated worsening of symptoms or of their condition, they were rescheduled for an immediate/timely in-person revisit with a spinal physician. (2) Fifty patients were offered telemedicine visits alone: 35 accepted: Next, from 2020 to 2021, we provided a questionnaire to 50 patients, we deemed eligible for teleconsultations: 35 agreed to the teleconsultations, while 15 refused (i.e., selecting direct in-person assessments). Results: We found a comparable quality for the clinical consultations/assessments provided in-person versus through telemedicine. Further, the additional 35 patients who were positively impressed with the quality of the medical care provided utilizing the telemedicine/remote visits alone. Conclusion: When pandemic conditions worsen, telemedicine appears to be a viable and important tool/ alternative for spinal surgeons to screen potential patients for treatment/surgery. This preliminary study suggests that a remote examination may be effective, particularly in patients who have previously undergone prior in-person evaluations.

Long versus short segment with intermediate screw fixation for burst fractures of thoracolumbar junction: radiological and clinical results.

BACKGROUND: Long segment fixation has been frequently used to treat thoracolumbar burst fractures. In our study we want to compare the long and short segment with intermediate screw fixation of thoracolumbar junction burst fractures in relation to radiological and clinical outcomes. METHODS: We collected the data of 115 patients, with thoracolumbar junction (T11-L2) burst fracture A3 or A4, according to AO classification. Patients were divided into two groups. Group A was treated by long segment fixation. Group B was treated by short segment fixation. At admission, after surgery, and at 12-month follow-up the patients were radiographically assessed for local kyphotic angle using the Cobb method. Patients were clinically evaluated with the Visual Analogue Scale (VAS) for back pain. RESULTS: The mean difference of the preoperative, immediate postoperative, and 12-month follow-up Cobb angle was significant in both groups (p = 0.018). The mean difference of the preoperative and immediate postoperative Cobb angle, Δ1, was significantly greater in group A than in group B (p = 0.038), indicating that the Cobb angle correction immediately postoperative was better in patients with double level fixation. The mean difference of the immediate postoperative and 12-month follow-up Cobb angle, Δ2, was significantly greater in group A than in group B (p = 0.007), indicating that the maintenance of local Cobb angle correction was better in patients with double level, long fixation. There was no difference in VAS values between Group B (single) and Group A (double) segment fixation immediately post operatively (p = 0.356) or after 12 months (p = 0.147). CONCLUSIONS: In A3 and A4 thoracolumbar junction fractures, long segment fixation can correct the local kyphosis Cobb angle and maintain the correction at 12-month follow-up better than short segment fixation with intermediate screws in the fractured vertebra. The radiological difference, however, was not predictive of clinical results.

Multidisciplinary Approach to Patients With Metastatic Spinal Cord Compression: A Diagnostic Therapeutic Algorithm to Improve the Neurological Outcome.

Introduction: The morbidity associated with metastatic spinal disease is significant because of spinal cord and/or nerve root compression. The purpose of this paper is to define a diagnostic-therapeutic path for patients with vertebral metastases and from this path to build an algorithm to reduce the devastating consequences of spinal cord compression. Materials and Methods: The algorithm is born from the experience of a primary care center. A spine surgeon, an emergency room (ER) physician, a neuroradiologist, a radiation oncologist, and an oncologist form the multidisciplinary team. The ER physician or the oncologist intercept the patient with symptoms and signs of a metastatic spinal cord compression. Once the suspicion is confirmed, the following steps of the flow-chart must be triggered. The spine surgeon takes charge of the patient and, on the base of the anamnestic data and neurological examination, defines the appropriate timing for magnetic resonance imaging (MRI) in collaboration with the neuroradiologist. From the MRI outcome, the spine surgeon and the radiation oncologist consult each other to define further therapeutic alternatives. If indicated, surgical treatment should precede radiation therapy. The oncologist gets involved after surgery for systemic therapy. Results: In 2021, the Spine and Spinal Cord Surgery department evaluated 257 patients with vertebral metastasis. Fifty-three patients presented with actual or incipient spinal cord compression. Among these, 27 were admitted due to rapid progression of symptoms, neurological deficits and/or spine instability signs. The level was thoracic in 21 cases, lumbar in 4 cases, cervical in 1 case, sacral in 1 case. Fifteen were operated on, 10 of these programmed and 5 in emergency. Discussion: Patients with a history of malignancy can present to the ER or to the oncology department with symptoms that must be correctly framed in the context of a metastatic involvement. Even when there is no previous cancer history, the patient's pain characteristics and clinical signs must be interpreted to yield the correct diagnosis of vertebral metastasis with incipient or current spinal cord compression. The awareness of the alert symptoms and the application of an integrated paradigm consent to frame the patients with spinal cord compression, obtaining the benefits of a homogeneous step-by-step diagnostic and therapeutic path. Early surgical or radiation therapy treatment gives the best hope for preventing the worsening, or even improving, the deficits. Conclusions: Metastatic spinal cord compression can cause neurological deficits compromising quality of life. Treatment strategies should be planned comprehensively. A multidisciplinary approach and the application of the proposed algorithm is of paramount importance to optimize the outcomes of these patients.

Spine Surgery in Italy in the COVID-19 Era: Proposal for Assessing and Responding to the Regional State of Emergency.

In December 2019, coronavirus disease 2019 (COVID-19) was discovered in Wuhan, Hubei province, from where it spread rapidly worldwide. COVID-19 characteristics (increased infectivity, rapid spread, and general population susceptibility) pose a great challenge to hospitals. Infectious disease, pulmonology, and intensive care units have been strengthened and expanded. All other specialties have been compelled to suspend or reduce clinical and elective surgical activities. The profound effects on spine surgery call for systematic approaches to optimizing the diagnosis and treatment of spinal diseases. Based on the experience of one Italian region, we draw an archetype for assessing the current and predicted level of stress in the health care system, with the aim of enabling hospitals to make better decisions during the pandemic. Further, we provide a framework that may help guide strategies for adapting surgical spine care to the conditions of epidemic surge.

Degenerative Lumbar Spine Stenosis Consensus Conference: the Italian job. Recommendations of the Spinal Section of the Italian Society of Neurosurgery.

In the modern era evidence-based medicine, guidelines and recommendations represent a key-point of daily activity. The Spinal Section of the Italian Society of Neurosurgery introduced some recommendations regarding Degenerative Lumbar Spine Stenosis based on those of the Spine Committee of World Federation of Neurosurgical Societies, revising them on the basis of Italian common practice. In June 2019, a Committee of 21 spine surgeons met in Rome to validate the recommendations of the WFNS. Furthermore, they decided to review the ones that did not reach a consensus to create Italian Recommendations on Degenerative Lumbar Spine Stenosis. A literature review of the last ten years was performed and the statements were voted using the Delphi method. Forty-one statements were discussed, and 7 statements were voted again to reach a consensus with respect to those of the WFNS. A total of 40 statements reached a consensus, of which 36 reached a positive consensus and 4 a negative consensus, while no consensus was reached in 1 case. Conservative multimodal therapy, tailored on the patient, is a reasonable and effective first option choice for the treatment of LSS patients with tolerable moderate symptoms. Surgical treatment is reserved for symptomatic patients non-responding to conservative treatment or with neurological deficits. The best surgical technique to use depends on personal experience; modern MISS techniques are equivalent to open decompressive surgery with some advantages and higher cost-effectiveness. Fusion surgery and mobility preserving surgery only have a marginal role in the treatment of DLSS without instability.

Deletion of a conserved Gata2 enhancer impairs haemogenic endothelium programming and adult Zebrafish haematopoiesis.

Gata2 is a key transcription factor required to generate Haematopoietic Stem and Progenitor Cells (HSPCs) from haemogenic endothelium (HE); misexpression of Gata2 leads to haematopoietic disorders. Here we deleted a conserved enhancer (i4 enhancer) driving pan-endothelial expression of the zebrafish gata2a and showed that Gata2a is required for HE programming by regulating expression of runx1 and of the second Gata2 orthologue, gata2b. By 5 days, homozygous gata2aΔi4/Δi4 larvae showed normal numbers of HSPCs, a recovery mediated by Notch signalling driving gata2b and runx1 expression in HE. However, gata2aΔi4/Δi4 adults showed oedema, susceptibility to infections and marrow hypo-cellularity, consistent with bone marrow failure found in GATA2 deficiency syndromes. Thus, gata2a expression driven by the i4 enhancer is required for correct HE programming in embryos and maintenance of steady-state haematopoietic stem cell output in the adult. These enhancer mutants will be useful in exploring further the pathophysiology of GATA2-related deficiencies in vivo.

Twelve-year results of LINAC-based radiosurgery for vestibular schwannomas.

PURPOSE: To report long-term outcomes of 53 patients with vestibular schwannomas (VS) submitted to a single high-dose LINAC-based radiosurgery (SRS) in our institution. METHODS: 48 (92%) patients were evaluable for clinical and MRI response as well as late toxicity. At a median follow-up of 12 years (range 2-16 years), local control (LC), hearing capacity, trigeminal and facial nerve function, and toxicity were assessed. Hearing capacity was classified according to the Gardner-Robertson scale, where class I-II patients had "serviceable hearing." RESULTS: Median dose of SRS was 16.5 Gy (range 13-20 Gy) and median tumor volume 1.7 cm3 (range 0.09-7.4 cm3). 35 (73%) patients were treated with SRS alone, in the remaining 13 (27%) patients, SRS was performed as salvage therapy for recurrent or progressive tumors after previous microsurgery. Before SRS, 44 patients (92%) had hearing loss and 25 (52%) had "non-serviceable" hearing. Tumor extension, classified with Koos categories, was grade I-II in 27 (56%) and grade III-IV in 21 (44%) cases. LC was 100% and hearing preservation in "serviceable hearing" patients was 91%. 4 (11%) patients developed incomplete and intermittent ipsilateral facial nerve palsy which regressed in a median time of 6 months. Trigeminal toxicity was registered in 11 (23%) patients, reversible in 6 (13%) and permanent in 5 (10%). Only Koos tumor grade III-IV significantly influenced late toxicity (p = 0.01). CONCLUSION: LC and hearing preservation after SRS were excellent. Toxicity proved acceptable. Although the median administered dose (16.5 Gy) was rather high, the only factor which significantly influenced late toxicity was Koos tumor grade III-IV.

LINAC-based radiosurgery for melanoma, sarcoma and renal cell carcinoma brain metastases.

BACKGROUND: The aim of this study was to report response, overall survival (OS) and toxicity in patients with radioresistant brain metastases (BM) treated with stereotactic radiosurgery (SRS). METHODS: Patients with renal cell carcinoma, melanoma and sarcoma with one to four brain metastases received SRS without whole brain radiotherapy. RESULTS: Fifty patients with 77 BM were treated. 46 (92%) patients with 71 BM were evaluable. Median follow-up was 67 months and median OS 11.8 months. At the time of analysis all patients had died. Brain control was conditioned by response to SRS (P<0.0001), while OS by histology (renal cell carcinoma versus melanoma and sarcoma) (P=0.04) and status of the tumour outside the brain (P=0.05). Treatment was well tolerated without more than grade 2 acute toxicity. CONCLUSIONS: Treatment of BM from radioresistant tumors with SRS assures good brain control and OS with low toxicity. Our data suggest a better prognosis associated to renal cell carcinoma histology.

Blood stem cell-forming haemogenic endothelium in zebrafish derives from arterial endothelium.

Haematopoietic stem cells are generated from the haemogenic endothelium (HE) located in the floor of the dorsal aorta (DA). Despite being integral to arteries, it is controversial whether HE and arterial endothelium share a common lineage. Here, we present a transgenic zebrafish runx1 reporter line to isolate HE and aortic roof endothelium (ARE)s, excluding non-aortic endothelium. Transcriptomic analysis of these populations identifies Runx1-regulated genes and shows that HE initially expresses arterial markers at similar levels to ARE. Furthermore, runx1 expression depends on prior arterial programming by the Notch ligand dll4. Runx1-/- mutants fail to downregulate arterial genes in the HE, which remains integrated within the DA, suggesting that Runx1 represses the pre-existing arterial programme in HE to allow progression towards the haematopoietic fate. These findings strongly suggest that, in zebrafish, aortic endothelium is a precursor to HE, with potential implications for pluripotent stem cell differentiation protocols for the generation of transplantable HSCs.

Etv6 activates vegfa expression through positive and negative transcriptional regulatory networks in Xenopus embryos.

VEGFA signaling controls physiological and pathological angiogenesis and hematopoiesis. Although many context-dependent signaling pathways downstream of VEGFA have been uncovered, vegfa transcriptional regulation in vivo remains unclear. Here, we show that the ETS transcription factor, Etv6, positively regulates vegfa expression during Xenopus blood stem cell development through multiple transcriptional inputs. In agreement with its established repressive functions, Etv6 directly inhibits expression of the repressor foxo3, to prevent Foxo3 from binding to and repressing the vegfa promoter. Etv6 also directly activates expression of the activator klf4; reflecting a genome-wide paucity in ETS-binding motifs in Etv6 genomic targets, Klf4 then recruits Etv6 to the vegfa promoter to activate its expression. These two mechanisms (double negative gate and feed-forward loop) are classic features of gene regulatory networks specifying cell fates. Thus, Etv6's dual function, as a transcriptional repressor and activator, controls a major signaling pathway involved in endothelial and blood development in vivo.

A community-based resource for automatic exome variant-calling and annotation in Mendelian disorders.

BACKGROUND: Mendelian disorders are mostly caused by single mutations in the DNA sequence of a gene, leading to a phenotype with pathologic consequences. Whole Exome Sequencing of patients can be a cost-effective alternative to standard genetic screenings to find causative mutations of genetic diseases, especially when the number of cases is limited. Analyzing exome sequencing data requires specific expertise, high computational resources and a reference variant database to identify pathogenic variants. RESULTS: We developed a database of variations collected from patients with Mendelian disorders, which is automatically populated thanks to an associated exome-sequencing pipeline. The pipeline is able to automatically identify, annotate and store insertions, deletions and mutations in the database. The resource is freely available online http://exome.tigem.it. The exome sequencing pipeline automates the analysis workflow (quality control and read trimming, mapping on reference genome, post-alignment processing, variation calling and annotation) using state-of-the-art software tools. The exome-sequencing pipeline has been designed to run on a computing cluster in order to analyse several samples simultaneously. The detected variants are annotated by the pipeline not only with the standard variant annotations (e.g. allele frequency in the general population, the predicted effect on gene product activity, etc.) but, more importantly, with allele frequencies across samples progressively collected in the database itself, stratified by Mendelian disorder. CONCLUSIONS: We aim at providing a resource for the genetic disease community to automatically analyse whole exome-sequencing samples with a standard and uniform analysis pipeline, thus collecting variant allele frequencies by disorder. This resource may become a valuable tool to help dissecting the genotype underlying the disease phenotype through an improved selection of putative patient-specific causative or phenotype-associated variations.

Neural stem cells and glioblastoma.

Glioblastoma multiforme represents one of the most common brain cancers with a rather heterogeneous cellular composition, as indicated by the term "multiforme". Recent reports have described the isolation and identification of cancer neural stem cells from human adult glioblastoma multiforme, which possess the capacity to establish, sustain, and expand these tumours, even under the challenging settings posed by serial transplantation experiments. Our study focused on the distribution of neural cancer stem cells inside the tumour. The study is divided into three phases: removal of tumoral specimens in different areas of the tumour (centre, periphery, marginal zone) in an operative room equipped with a 1.5 T scanner; isolation and characterization of neural cancer stem cells from human adult glioblastoma multiforme; identification of neural cancer stem cell distribution inside the tumour.

Mantra 2.0: an online collaborative resource for drug mode of action and repurposing by network analysis.

SUMMARY: Elucidation of molecular targets of a compound [mode of action (MoA)] and its off-targets is a crucial step in drug development. We developed an online collaborative resource (MANTRA 2.0) that supports this process by exploiting similarities between drug-induced transcriptional profiles. Drugs are organized in a network of nodes (drugs) and edges (similarities) highlighting 'communities' of drugs sharing a similar MoA. A user can upload gene expression profiles before and after drug treatment in one or multiple cell types. An automated processing pipeline transforms the gene expression profiles into a unique drug 'node' embedded in the drug-network. Visual inspection of the neighbouring drugs and communities helps in revealing its MoA and to suggest new applications of known drugs (drug repurposing). MANTRA 2.0 allows storing and sharing user-generated network nodes, thus making MANTRA 2.0 a collaborative ever-growing resource. AVAILABILITY AND IMPLEMENTATION: The web tool is freely available for academic use at http://mantra.tigem.it.