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Cranial vault remodelling for craniosynostosis aims to increase intracranial volume to facilitate brain growth, avoid the development of raised intracranial pressure and address cosmesis. The extent of vault expansion is predominantly limited by scalp closure and reconstruction technique. Virtual surgical planning tools have been developed to predict post-operative changes and guide expansion. We present a validation study of a novel 'Dura-based Automated Vault Expansion-Remodeling' (DAVE-R) model to guide pre-operative planning for fronto-orbital advancement and remodelling (FOAR)., Methods: Patients with trigonocephaly who underwent FOAR with pre- and post-operative imaging from 2018 to 2020 were identified from a prospectively maintained database. Post-operative scans, normative atlas and whole brain parcellation were registered to the pre-operative images to quantify the change in intracranial volume and morphology (utilising measurement of fronto-orbital advancement and bifrontozygomatic distance) compared to that predicted by the DAVE-R model., Results: Ten patients were included. The DAVE-R model predicted bifrontozygomatic distances of 92.0 + / - 5.14 mm (mean + /SD), which closely matched the post-operative results of 92.7 + / - 6.02 mm (mean + / - SD); (t(d.f. 9) = -0.306, p = 0.77). The fronto-orbital advancement predicted by the DAVE-R method was 11.5 + / - 1.96 mm (mean + / - SD) which was significantly greater than 8.6 + / - 2.94 mm (mean ± SD); (t(d.f. 9) = 3.137, p = 0.01) achieved post-operatively., Conclusions: We demonstrate that the DAVE-R model provides an objective means of extracting realistic surgical goals in patients undergoing FOAR for trigonocephaly that closely correlates with post-operative outcomes. The normative dural model warrants further study and validation for other forms of craniosynostosis correction., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The Second International Conference of the World Society for Virology (WSV), hosted by Riga Stradiņš University, was held in Riga, Latvia, on June 15-17th, 2023. It prominently highlighted the recent advancements in different disciplines of virology. The conference had fourteen keynote speakers covering diverse topics, including emerging virus pseudotypes, Zika virus vaccine development, herpesvirus capsid mobility, parvovirus invasion strategies, influenza in animals and birds, West Nile virus and Marburg virus ecology, as well as the latest update in animal vaccines. Discussions further explored SARS-CoV-2 RNA replicons as vaccine candidates, SARS-CoV-2 in humans and animals, and the significance of plant viruses in the 'One Health' paradigm. The presence of the presidents from three virology societies, namely the American, Indian, and Korean Societies for Virology, highlighted the event's significance. Additionally, past president of the American Society for Virology (ASV), formally declared the partnership between ASV and WSV during the conference., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: The piriform cortex (PC) occupies both banks of the endorhinal sulcus and has an important role in the pathophysiology of temporal lobe epilepsy (TLE). A recent study showed that resection of more than 50% of PC increased the odds of becoming seizure free by a factor of 16., Objective: We report the feasibility of manual segmentation of PC and application of the Geodesic Information Flows (GIF) algorithm to automated segmentation, to guide resection., Methods: Manual segmentation of PC was performed by two blinded independent examiners in 60 patients with TLE (55% Left TLE, 52% female) with a median age of 35 years (IQR, 29-47 years) and 20 controls (60% Women) with a median age of 39.5 years (IQR, 31-49). The GIF algorithm was used to create an automated pipeline for parcellating PC which was used to guide excision as part of temporal lobe resection for TLE., Results: Right PC was larger in patients and controls. Parcellation of PC was used to guide anterior temporal lobe resection, with subsequent seizure freedom and no visual field or language deficit., Conclusion: Reliable segmentation of PC is feasible and can be applied prospectively to guide neurosurgical resection that increases the chances of a good outcome from temporal lobe resection for TLE.

Infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are rhabdoviruses in two different species belonging to the Novirhabdovirus genus. IHNV has a narrow host range restricted to trout and salmon species, and viruses in the M genogroup of IHNV have high virulence in rainbow trout ( Oncorhynchus mykiss ). In contrast, the VHSV genotype IVb that invaded the Great Lakes in the United States has a broad host range, with high virulence in yellow perch ( Perca flavescens ), but not in rainbow trout. By using reverse-genetic systems of IHNV-M and VHSV-IVb strains, we generated six IHNV:VHSV chimeric viruses in which the glycoprotein (G), non-virion-protein (NV), or both G and NV genes of IHNV-M were replaced with the analogous genes from VHSV-IVb, and vice versa. These chimeric viruses were used to challenge groups of rainbow trout and yellow perch. The parental recombinants rIHNV-M and rVHSV-IVb were highly virulent in rainbow trout and yellow perch, respectively. Parental rIHNV-M was avirulent in yellow perch, and chimeric rIHNV carrying G, NV, or G and NV genes from VHSV-IVb remained low in virulence in yellow perch. Similarly, the parental rVHSV-IVb exhibited low virulence in rainbow trout, and chimeric rVHSV with substituted G, NV, or G and NV genes from IHNV-M remained avirulent in rainbow trout. Thus, the G and NV genes of either virus were not sufficient to confer high host-specific virulence when exchanged into a heterologous species genome. Some exchanges of G and/or NV genes caused a loss of host-specific virulence, providing insights into possible roles in viral virulence or fitness, and interactions between viral proteins.

Citrobacter freundii, a common pathogen of freshwater fish, causes significant commercial losses to the global fish farming industry. In the present study, a highly pathogenic C. freundii strain was isolated and identified from largemouth bass (Micropterus salmoides). The pathogenicity and antibiotic sensitivity of the C. freundii strain were evaluated, and the histopathology and host immune response of largemouth bass infected with C. freundii were investigated. The results showed that C. freundii was the pathogen causing disease outbreaks in largemouth bass, and the infected fish showed typical signs of acute hemorrhages and visceral enlargement. Antimicrobial susceptibility testing showed that the C. freundii strain was resistant to Kanamycin, Medimycin, Clindamycin, Penicillin, Oxacillin, Ampicillin, Cephalexin, Cefazolin, Cefradine and Vancomycin. Histopathological analysis showed different pathological changes in major tissues of diseased fish. In addition, humoral immune factors such as superoxide dismutase (SOD), catalase (CAT) and lysozyme (LZM) were used as serum indicators to evaluate the immune response of largemouth bass after infection. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression pattern of immune-related genes (CXCR1, IL-8, IRF7, IgM, CD40, IFN-γ, IL-1β, Hep1, and Hep2) in liver, spleen, and head kidney tissues, which demonstrated a strong immune response induced by C. freundii infection in largemouth bass. The present study provides insights into the pathogenic mechanism of C. freundii and immune response in largemouth bass, promoting the prevention and treatment of diseases caused by C. freundii infection., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objective: Encephaloceles are considered to result from defects in the developing skull through which meninges, and potentially brain tissue, herniate. The pathological mechanism underlying this process is incompletely understood. We aimed to describe the location of encephaloceles through the generation of a group atlas to determine whether they occur at random sites or clusters within distinct anatomical regions., Methods: Patients diagnosed with cranial encephaloceles or meningoceles were identified from a prospectively maintained database between 1984 and 2021. Images were transformed to atlas space using non-linear registration. The bone defect, encephalocele and herniated brain contents were manually segmented allowing for a 3-dimensional heat map of encephalocele locations to be generated. The centroids of the bone defects were clustered utilising a K-mean clustering machine learning algorithm in which the elbow method was used to identify the optimal number of clusters., Results: Of the 124 patients identified, 55 had volumetric imaging in the form of MRI (48/55) or CT (7/55) that could be used for atlas generation. Median encephalocele volume was 14,704 (IQR 3655-86,746) mm 3 and the median surface area of the skull defect was 679 (IQR 374-765) mm 2 . Brain herniation into the encephalocele was found in 45% (25/55) with a median volume of 7433 (IQR 3123-14,237) mm 3 . Application of the elbow method revealed 3 discrete clusters: (1) anterior skull base (22%; 12/55), (2) parieto-occipital junction (45%; 25/55) and (3) peri-torcular (33%; 18/55). Cluster analysis revealed no correlation between the location of the encephalocele with gender (χ 2 (2, n = 91) = 3.86, p = 0.15). Compared to expected population frequencies, encephaloceles were relatively more common in Black, Asian and Other compared to White ethnicities. A falcine sinus was identified in 51% (28/55) of cases. Falcine sinuses were more common (χ 2 (2, n = 55) = 6.09, p = 0.05) whilst brain herniation was less common (χ 2 (2, n = 55) = .16.24, p < 0.0003) in the parieto-occipital location., Conclusion: This analysis revealed three predominant clusters for the location of encephaloceles, with the parieto-occipital junction being the most common. The stereotypic location of encephaloceles into anatomically distinct clusters and the coexistence of distinct venous malformations at certain sites suggests that their location is not random and raises the possibility of distinct pathogenic mechanisms unique to each of these regions., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The largemouth bass virus (LMBV) isolate of Santee-Cooper ranavirus showed evidence of widespread infection in adult fish, but disease presentation caused by different viral strains exhibited considerable difference. In this study, a highly pathogenic LMBV-like resembling Santee-Cooper ranavirus was isolated and identified from juvenile largemouth bass. The pathogenicity and dynamic distribution of LMBV-like strain, histopathological analysis and host immune response of juvenile largemouth bass infected with LMBV-like were investigated. The results show that LMBV-like was highly pathogenic to juvenile fish, and the infected fish showed typical signs of acute haemorrhages and visceral enlargement. LMBV-like positive cells were found in the liver, spleen, kidney, gills, and intestinal tissue, and the virus content in spleen was the highest. Histopathological analysis showed different pathological changes in major tissues of diseased fish, mostly manifested as infiltration of inflammatory cell and histiocyte necrosis. In addition, humoral immune factors such as superoxide dismutase (SOD), catalase (CAT) and acid phosphatase (ACP) were used as serum indicators to evaluate the immune response of juvenile fish after infection. Quantitative real-time PCR (qRT-PCR) was used to evaluate the expression patterns of immune-related genes (CD40, IFN-γ, IgM, IL-1β, IL-8, IL-12a, Mxd3, TGF-β, and TNFα) in liver, spleen, and head kidney tissues. The results showed that immunological activity of the juvenile largemouth bass was significantly enhanced by LMBV-like infection. This research comprehensively systematically revealed the pathogenic characteristics of LMBV-like separated from juvenile largemouth bass and properties of the host's immune response caused by the virus infection, which providing a basis for further exploring the interaction between the virus and the host, and prevention and treatment of disease caused by Santee-Cooper ranavirus., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Salmonid novirhabdovirus (IHNV) causes infectious haematopoietic necrosis (IHN) in salmonid species. Despite an injectable plasmid-based DNA vaccine of the glycoprotein (G) gene is effective, there are no oral vaccines for mass vaccination of rainbow trout ( Oncorhynchus mykiss ) fry. Recombinant baculoviruses were generated, used in cabbage looper ( Trichoplusia ni ) insect larvae to produce IHNV G and IHNV G-C5a proteins. Western blotting and chemiluminescence assays confirmed the expression of recombinant proteins, which were added to the fish feeding and top-coated with unflavored gelatin binder. Commercial rainbow trout were fed with experimental diets containing either IHNV G or IHNV G-C5a proteins for 2 weeks, and boosted 4 weeks after. Four weeks post-booster, fish were challenged with IHNV by immersion. Survival upon the infection challenge was evaluated. Spleen were sampled at 7 and 14 days post infection (dpi). Non-vaccinated and IHNV G fed trout reached a mortality of 91.7 and 97.6%, and 70.9 and 88.4%, respectively at 8 and 15 dpi. The IHNV G-C5a fed group exhibited a reduced mortality of 51.2% at 8 dpi, reaching 81.7% at 15 dpi, suggesting some level of antiviral protection. The individual viral load was measured by RT-qPCR detection of IHNV N gene, showing no significant difference across experimental groups. The transcription modulation of selected immune response markers was evaluated across experimental groups, including Type I IFN-a, Mx-1, CD4, and IgM. Further study is needed to assess how new oral vaccines may become effective to mitigate IHNV pathogenesis in juvenile trout by modulating the host immune response to protect towards IHNV exposure., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Interferon-stimulated gene 15 (ISG15) regulates cellular life processes, including defense responses against infection by a variety of viral pathogens, by binding to target proteins. At present, various fish ISG15s have been identified, but the biological function of ISG15 in snakehead fish is still unclear. In this study, total RNA was extracted from snakehead fish cell line E11, ds cDNA was synthesized and purified using SMART technology, and the resulting cDNA library was screened by co-transforming yeast cells. The library titer was 4.28 × 10 9  CFU/mL. Using snakehead ISG15 as the bait protein, the recombinant bait vector pGBKT7-ISG15 was constructed and transformed into the yeast strain Y2HGold. The toxicity and self-activation activity of the bait vector were detected on the deficient medium, and the prey proteins interacting with ISG15 were screened. In total, 19 interacting proteins of ISG15 were identified, including mitotic checkpoint protein BUB3, hypothetical protein SnRVgp6, elongation factor 1-beta, 60S ribosomal protein L9, dual specificity protein phosphatase 5-like, eukaryotic translation initiation factor 3 subunit I and ferritin. A yeast spotting assay further probed the interaction between ISG15 and DUSP5. These results increase our understanding of the interaction network of snakehead ISG15 and will aid in exploring the underlying mechanisms of snakehead ISG15 functions in the future., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The coronavirus disease-19 (COVID-19) pandemic has already claimed millions of lives and remains one of the major catastrophes in the recorded history. While mitigation and control strategies provide short term solutions, vaccines play critical roles in long term control of the disease. Recent emergence of potentially vaccine-resistant and novel variants necessitated testing and deployment of novel technologies that are safe, effective, stable, easy to administer, and inexpensive to produce. Here we developed three recombinant Newcastle disease virus (rNDV) vectored vaccines and assessed their immunogenicity, safety, and protective efficacy against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in mice and hamsters. Intranasal administration of rNDV-based vaccine candidates elicited high levels of neutralizing antibodies. Importantly, the nasally administrated vaccine prevented lung damage, and significantly reduced viral load in the respiratory tract of vaccinated animal which was compounded by profound humoral immune responses. Taken together, the presented NDV-based vaccine candidates fully protected animals against SARS-CoV-2 challenge and warrants evaluation in a Phase I human clinical trial as a promising tool in the fight against COVID-19., (© 2022. The Author(s).)

Avian influenza virus (AIV) represents a major concern with productive implications in poultry systems but it is also a zoonotic agent that possesses an intrinsic pandemic risk. AIV is an enveloped, negative-sense and single-stranded RNA virus with a segmented genome. The eight genomic segments, comprising the whole genome, encode for eleven proteins. Within these proteins, Hemagglutinin (HA) and Neuraminidase (NA) are the most relevant for studies of evolution and pathogenesis considering their role in viral replication, and have also been used for classification purposes. Migratory birds are the main hosts and play a pivotal role in viral evolution and dissemination due to their migratory routes that comprise large regions worldwide. Altogether, viral and reservoir factors contribute to the emergence of avian influenza viruses with novel features and pathogenic potentials. The study aimed to conduct surveillance of AIVs in wild birds from Peru. A multi-site screening of feces of migratory birds was performed to isolate viruses and to characterize the whole genome sequences, especially the genes coding for HA and NA proteins. Four-hundred-twenty-one (421) fecal samples, collected between March 2019 and March 2020 in Lima, were obtained from 21 species of wild birds. From these, we isolated five AIV from whimbrel, kelp gull, Franklin's gulls and Mallard, which were of low pathogenicity, including four subtypes as H6N8, H13N6, H6N2 and H2N6. Genetic analysis of HA and NA genes revealed novel features in these viruses and phylogenetic analysis exhibited a close relationship with those identified in North America (US and Canada). Furthermore, H2N6 isolate presented a NA sequence with higher genetic relationship to Chilean isolates. These results highlight that the geographical factor is of major relevance in the evolution of AIV, suggesting that AIV circulating in Peru might represent a new site for the emergence of reassortant AIVs., Competing Interests: The authors have declared that no competing interests exist.

Objective: Accurate preoperative predictions of seizure freedom following surgery for focal drug resistant epilepsy remain elusive. Our objective was to systematically evaluate all meta-analyses of epilepsy surgery with seizure freedom as the primary outcome, to identify clinical features that are consistently prognostic and should be included in the future models., Methods: We searched PubMed and Cochrane using free-text and Medical Subject Heading (MeSH) terms according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses. This study was registered on PROSPERO. We classified features as prognostic, non-prognostic and uncertain and into seven subcategories: 'clinical', 'imaging', 'neurophysiology', 'multimodal concordance', 'genetic', 'surgical technique' and 'pathology'. We propose a structural causal model based on these features., Results: We found 46 features from 38 meta-analyses over 22 years. The following were consistently prognostic across meta-analyses: febrile convulsions, hippocampal sclerosis, focal abnormal MRI, Single-Photon Emission Computed Tomography (SPECT) coregistered to MRI, focal ictal/interictal EEG, EEG-MRI concordance, temporal lobe resections, complete excision, histopathological lesions, tumours and focal cortical dysplasia type IIb. Severe learning disability was predictive of poor prognosis. Others, including sex and side of resection, were non-prognostic. There were limited meta-analyses investigating genetic contributions, structural connectivity or multimodal concordance and few adjusted for known confounders or performed corrections for multiple comparisons., Significance: Seizure-free outcomes have not improved over decades of epilepsy surgery and despite a multitude of models, none prognosticate accurately. Our list of multimodal population-invariant prognostic features and proposed structural causal model may serve as an objective foundation for statistical adjustments of plausible confounders for use in high-dimensional models., Prospero Registration Number: CRD42021185232., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY. Published by BMJ.)

This communication summarizes the presentations given at the 1st international conference of the World Society for Virology (WSV) held virtually during 16-18 June 2021, under the theme of tackling global viral epidemics. The purpose of this biennial meeting is to foster international collaborations and address important viral epidemics in different hosts. The first day included two sessions exclusively on SARS-CoV-2 and COVID-19. The other two days included one plenary and three parallel sessions each. Last not least, 16 sessions covered 140 on-demand submitted talks. In total, 270 scientists from 49 countries attended the meeting, including 40 invited keynote speakers., (Copyright © 2021 Elsevier Inc. All rights reserved.)

White matter bundle segmentation using diffusion MRI fiber tractography has become the method of choice to identify white matter fiber pathways in vivo in human brains. However, like other analyses of complex data, there is considerable variability in segmentation protocols and techniques. This can result in different reconstructions of the same intended white matter pathways, which directly affects tractography results, quantification, and interpretation. In this study, we aim to evaluate and quantify the variability that arises from different protocols for bundle segmentation. Through an open call to users of fiber tractography, including anatomists, clinicians, and algorithm developers, 42 independent teams were given processed sets of human whole-brain streamlines and asked to segment 14 white matter fascicles on six subjects. In total, we received 57 different bundle segmentation protocols, which enabled detailed volume-based and streamline-based analyses of agreement and disagreement among protocols for each fiber pathway. Results show that even when given the exact same sets of underlying streamlines, the variability across protocols for bundle segmentation is greater than all other sources of variability in the virtual dissection process, including variability within protocols and variability across subjects. In order to foster the use of tractography bundle dissection in routine clinical settings, and as a fundamental analytical tool, future endeavors must aim to resolve and reduce this heterogeneity. Although external validation is needed to verify the anatomical accuracy of bundle dissections, reducing heterogeneity is a step towards reproducible research and may be achieved through the use of standard nomenclature and definitions of white matter bundles and well-chosen constraints and decisions in the dissection process., (Copyright © 2021. Published by Elsevier Inc.)

Newcastle disease virus (NDV) strain R2B, with an altered fusion protein cleavage site, was used as a viral vector to deliver the immunogenic genes VP2 and VP1 of chicken infectious anaemia virus (CIAV) to generate a bivalent vaccine candidate against these diseases in chickens. The immunogenic genes of CIAV were expressed as a single transcriptional unit from the NDV backbone and the two CIA viral proteins were obtained as separate entities using a self-cleaving foot-and-mouth disease virus 2A protease sequence between them. The recombinant virus (rR2B-FPCS-CAV) had similar growth kinetics as that of the parent recombinant virus (rR2B-FPCS) in vitro with similar pathogenicity characteristics. The bivalent vaccine candidate when given in specific pathogen-free chickens as primary and booster doses was able to elicit robust humoral and cell-mediated immune (CMI) responses obtained in a vaccination study that was conducted over a period of 15 weeks. In an NDV and CIAV ELISA trial, there was a significant difference in the titres of antibody between vaccinated and control groups which showed slight reduction in antibody titre by 56 days of age. Hence, a second booster was administered and the antibody titres were maintained until 84 days of age. Similar trends were noticed in CMI response carried out by lymphocyte transformation test, CD4 + and CD8 + response by flow cytometry analysis and response of real time PCR analysis of cytokine genes. Birds were challenged with virulent NDV and CIAV at 84 days and there was significant reduction in the NDV shed on the 2nd and 4th days post challenge in vaccinated birds as compared to unvaccinated controls. Haematological parameters comprising PCV, TLC, PLC and PHC were estimated in birds that were challenged with CIAV that indicated a significant reduction in the blood parameters of controls. Our findings support the development and assessment of a bivalent vaccine candidate against NDV and CIAV in chickens.

Laser interstitial thermal therapy (LiTT) isa minimally invasive alternative to conventional open surgery for drug-resistant focal mesial temporal lobe epilepsy (MTLE). Recent studies suggest that higher seizure freedom rates are correlated with maximal ablation of the mesialhippocampal head, whilst sparing of the parahippocampal gyrus (PHG) may reduce neuropsychological sequelae. Current commercially available laser catheters are inserted following manually planned straight-line trajectories, which cannot conform to curved brain structures, such as the hippocampus, without causing collateral damage or requiring multiple insertions., Objectives: The clinical feasibility and potential of curved LiTT trajectories through steerable needles has yet to be investigated. This is the focus of our work., Methods: We propose a GPU-accelerated computer-assisted planning (CAP) algorithm for steerable needle insertions that generates optimized curved 3D trajectories with maximal ablation of the amygdalohippocampal complex and minimal collateral damage to nearby structures, while accounting for a variable ablation diameter ( 5-15 mm)., Results: Simulated trajectories and ablations were performed on 5 patients with mesial temporal sclerosis (MTS), which were identified from a prospectively managed database. The algorithm generated obstacle-free paths with significantly greater target area ablation coverage and lower PHG ablation variance compared to straight line trajectories., Conclusions: The presented CAP algorithm returns increased ablation of the amygdalohippocampal complex, with lower patient risk scores compared to straight-line trajectories., Significance: This is the first clinical application of preoperative planning for steerable needle based LiTT. This study suggests that steerableneedles have the potential to improve LiTT procedure efficacy whilst improving the safety and should thus be investigated further.

Recombinant Newcastle disease virus vectors have gained a lot of interest for its oncolytic virus therapy and cancer immune therapeutic properties due to its selective replication to high titers in cancer cells. The aim of this study was to find out the oncolytic effects of mesogenic recombinant NDV strain R2B-GFP on murine mammary tumor cell line 4T1 and murine melanoma cell line B16-F10. The anti-tumor effects of R2B-GFP virus were studied via expression of virus transgene GFP in cancer cells, evaluating its cytotoxicity and cell migration efficacies by MTT and wound healing assays respectively. In addition, the underlying apoptotic mechanism of R2B-GFP virus was estimated by TUNEL assay, colorimetric estimation of Caspase-3, 8 and 9 and the estimation of Bax to Bcl-2 ratio. The results showed a significant decrease in viability of both 4T1 and B16-F10 cells infected with R2B-GFP virus at 0.1 and 1 MOI. R2B-GFP virus could significantly induce apoptosis in the 4T1 and B16-F10 cells as compared to the uninfected control. Further, a flow cytometry analysis on apoptotic cells percentage and mitochondria membrane permeability test was also studied in R2B-GFP virus treated 4T1 and B16-F10 cell lines. The R2B-GFP virus caused an increase in loss of mitochondrial membrane permeability in both 4T1 and B16-F10 cells indicating the involvement of mitochondrial regulated cell death. Thus, the recombinant virus R2B-GFP virus proved to be a valid candidate for oncolytic viral therapy in 4T1 and B16-F10 cells., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

There has been a significant rise in robotic trajectory guidance devices that have been utilised for stereotactic neurosurgical procedures. These devices have significant costs and associated learning curves. Previous studies reporting devices usage have not undertaken prospective parallel-group comparisons before their introduction, so the comparative differences are unknown. We study the difference in stereoelectroencephalography electrode implantation time between a robotic trajectory guidance device (iSYS1) and manual frameless implantation (PAD) in patients with drug-refractory focal epilepsy through a single-blinded randomised control parallel-group investigation of SEEG electrode implantation, concordant with CONSORT statement. Thirty-two patients (18 male) completed the trial. The iSYS1 returned significantly shorter median operative time for intracranial bolt insertion, 6.36 min (95% CI 5.72-7.07) versus 9.06 min (95% CI 8.16-10.06), p = 0.0001. The PAD group had a better median target point accuracy 1.58 mm (95% CI 1.38-1.82) versus 1.16 mm (95% CI 1.01-1.33), p = 0.004. The mean electrode implantation angle error was 2.13° for the iSYS1 group and 1.71° for the PAD groups (p = 0.023). There was no statistically significant difference for any other outcome. Health policy and hospital commissioners should consider these differences in the context of the opportunity cost of introducing robotic devices.Trial registration: ISRCTN17209025 ( https://doi.org/10.1186/ISRCTN17209025 )., (© 2021. The Author(s).)

Purpose: Instability of the craniocervical junction in paediatric patients with skeletal dysplasia poses a unique set of challenges including anatomical abnormalities, poor bone quality, skeletal immaturity and associated general anaesthetic risks. Instrumented fixation provides optimal stabilisation and fusion rates. The small vertebrae make the placement of C2 pedicle screws technically demanding with low margins of error between the spinal canal and the vertebral artery., Methods: We describe a novel clinical strategy utilising 3D-printed spinal screw trajectory guides (3D-SSTG) for individually planned C2 pedicle and laminar screws. The technique is based on a pre-operative CT scan and does not require intraoperative CT imaging. This reduces the radiation burden to the patient and forgoes the associated time and cost. The time for model generation and sterilisation was < 24 h., Results: We describe two patients (3 and 6 years old) requiring occipitocervical instrumented fixation for cervical myelopathy secondary to Morquio syndrome with 3D-SSTGs. In the second case, bilateral laminar screw trajectories were also incorporated into the same guide due to the presence of high-riding vertebral arteries. Registration of the postoperative CT to the pre-operative imaging revealed that screws were optimally placed and accurately followed the predefined trajectory., Conclusion: To our knowledge, we present the first clinical report of 3D-printed spinal screw trajectory guides at the craniocervical junction in paediatric patients with skeletal dysplasia. The novel combination of multiple trajectories within the same guide provides the intraoperative flexibility of potential bailout options. Future studies will better define the potential of this technology to optimise personalised non-standard screw trajectories., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objective: Anteromesial temporal lobe resection (ATLR) results in long-term seizure freedom in patients with drug-resistant focal mesial temporal lobe epilepsy (MTLE). There is significant anatomical variation in the anterior projection of the optic radiation (OR), known as Meyer's loop, between individuals and between hemispheres in the same individual. Damage to the OR results in contralateral superior temporal quadrantanopia that may preclude driving in 33%-66% of patients who achieve seizure freedom. Tractography of the OR has been shown to prevent visual field deficit (VFD) when surgery is performed in an interventional MRI (iMRI) suite. Because access to iMRI is limited at most centers, the authors investigated whether use of a neuronavigation system with a microscope overlay in a conventional theater is sufficient to prevent significant VFD during ATLR., Methods: Twenty patients with drug-resistant MTLE who underwent ATLR (9 underwent right-side ATLR, and 9 were male) were recruited to participate in this single-center prospective cohort study. Tractography of the OR was performed with preoperative 3-T multishell diffusion data that were overlaid onto the surgical field by using a conventional neuronavigation system linked to a surgical microscope. Phantom testing confirmed overlay projection errors of < 1 mm. VFD was quantified preoperatively and 3 to 12 months postoperatively by using Humphrey and Esterman perimetry., Results: Perimetry results were available for all patients postoperatively, but for only 11/20 (55%) patients preoperatively. In 1/20 (5%) patients, a significant VFD occurred that would prevent driving in the UK on the basis of the results on Esterman perimetry. The VFD was identified early in the series, despite the surgical approach not transgressing OR tractography, and was subsequently found to be due to retraction injury. Tractography was also used from this point onward to inform retractor placement, and no further significant VFDs occurred., Conclusions: Use of OR tractography with overlay outside of an iMRI suite, with application of an appropriate error margin, can be used during approach to the temporal horn of the lateral ventricle and carries a 5% risk of VFD that is significant enough to preclude driving postoperatively. OR tractography can also be used to inform retractor placement. These results warrant a larger prospective comparative study of the use of OR tractography-guided mesial temporal resection.

Introduction: The COVID-19 pandemic had an unprecedented global socioeconomic impact. Responses to pandemics include strategies to accumulate vast stockpiles of vital medical equipment. In such times of desperation, 3D-printing could be a life-saving alternative., Methods: We undertook a PRISMA systematic review of 3D printing solutions in response to COVID-19 utilising the PICO methodology. The objectives were to identify the uses of 3D printing during the COVID-19 pandemic, determine the extent of preclinical testing, comparison to commercial alternatives, presence of regulatory approvals and replicability regarding the description of the printing parameters and the availability of the print file., Results: Literature searches of MEDLINE (OVID interface)/ PubMed identified 601 studies. Of these, 10 studies fulfilled the inclusion and exclusion criteria. Reported uses of 3D printing included personal protective equipment (PPE), nasopharyngeal swabs and adjunctive anaesthetic equipment. Few studies undertook formal safety and efficacy testing before clinical use with only one study comparing to the commercial equivalent. Six articles made their model print files available for wider use., Conclusion: We describe a protocol for a systematic review of 3D-printed healthcare solutions in response to COVID-19. This remains a viable method of producing vital healthcare equipment when supply chains are exhausted. We hope that this will serve as a summary of innovative 3D-printed solutions during the peak of the pandemic and also highlight concerns and omissions regarding safety and efficacy testing that should be addressed urgently in preparation for a subsequent resurgences and future pandemics ., Competing Interests: All authors declare: no support from any organisation or financial relationships with any organisations that might have an interest in the submitted work., (© 2021 The Authors.)

A new grass carp reovirus (GCRV), healthy grass carp reovirus (HGCRV), was isolated from grass carp in 2019. Its complete genome sequence was determined and contained 11 dsRNAs with a total size of 23,688 bp and 57.2 mol% G+C content, encoding 12 proteins. All segments had conserved 5' and 3' termini. Sequence comparisons showed that HGCRV was closely related to GCRV-873 (GCRV-I; 69.57-96.71% protein sequence identity) but shared only 22.65-45.85% and 23.37-43.39% identities with GCRV-HZ08 and Hubei grass carp disease reovirus (HGDRV), respectively. RNA-dependent RNA-polymerase (RdRp) protein-based phylogenetic analysis showed that HGCRV clustered with Aquareovirus -C ( AqRV -C) prior to joining a branch common with other aquareoviruses. Further analysis using VP6 amino acid sequences from Chinese GCRV strains showed that HGCRV was in the same evolutionary cluster as GCRV-I. Thus, HGCRV could be a new GCRV isolate of GCRV-I but is distantly related to other known GCRVs. Grass carp infected with HGCRV did not exhibit signs of hemorrhage. Interestingly, the isolate induced a typical cytopathic effect in fish cell lines, such as infected cell shrank, apoptosis, and plague-like syncytia. Further analysis showed that HGCRV could proliferate in grass carp liver (L28824), gibel carp brain (GiCB), and other fish cell lines, reaching a titer of up to 7.5 × 10 4 copies/μL.

Purpose of Review: Surgery can provide a robust long-standing seizure remission in drug-refractory mesial temporal lobe epilepsy (MTLE). Despite this, a significant proportion of postoperative patients are ineligible to gain a driving licence due to the size of the subsequent visual field defect (VFD). The amygdala and hippocampus are intimately related to several important white fibre association tracts and damage to the optic radiation results in a contralateral superior quadrantanopia. For this reason, several different modifications to established surgical approaches and novel techniques have recently been applied to mitigate or prevent damage to the optic radiation. There is still no consensus on which operative technique results in optimal outcomes regarding seizure remission, neuropsychological sequelae and VFD rates. We explore contemporary surgical approaches to the mesial temporal lobe and describe the intraoperative use of tractography and iMRI in preventing VFDs., Recent Findings: Established approaches for the surgical treatment of MTLE include standardized approaches in the form of anterior temporal lobectomies, selective approaches and various modifications thereof. Recent advancements in microsurgical techniques have seen numerous modifications to these approaches to spare the optic radiation as well as the introduction of minimally invasive alternatives such as laser interstitial thermal therapy (LITT) and stereotactic radiosurgery (SRS). The intraoperative use of optic radiation tractography through overlays in the operative microscope and interventional MRI suites to correct for brain shift have been shown to reduce VFDs., Summary: VFDs following the surgical treatment of drug-refractory MTLE can have a significant impact on the quality of life. Each of the surgical techniques carries a risk to the visual pathways but the use of minimally invasive techniques as well as surgical adjuncts may reduce or prevent acquired VFDs., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

The grass carp reovirus (GCRV) causes severe hemorrhagic disease with high mortality and leads to serious economic losses in the grass carp ( Ctenopharyngodon idella ) industry in China. Oral vaccine has been proven to be an effective method to provide protection against fish viruses. In this study, a recombinant baculovirus BmNPV-VP35-VP4 was generated to express VP35 and VP4 proteins from GCRV type Ⅱ via Bac-to-Bac baculovirus expression system. The expression of recombinant VP35-VP4 protein (rVP35-VP4) in Bombyx mori embryo cells (BmE) and silkworm pupae was confirmed by Western blotting and immunofluorescence assay (IFA) after infection with BmNPV-VP35-VP4. To vaccinate the grass carp by oral route, the silkworm pupae expressing the rVP35-VP4 proteins were converted into a powder after freeze-drying, added to artificial feed at 5% and fed to grass carp (18 ± 1.5 g) for six weeks, and the immune response and protective efficacy in grass carp after oral vaccination trial was thoroughly investigated. This included blood cell counting and classification, serum antibody titer detection, immune-related gene expression and the relative percent survival rate in immunized grass carp. The results of blood cell counts show that the number of white blood cells in the peripheral blood of immunized grass carp increased significantly from 14 to 28 days post-immunization (dpi). The differential leukocyte count of neutrophils and monocytes were significantly higher than those in the control group at 14 dpi. Additionally, the number of lymphocytes increased significantly and reached a peak at 28 dpi. The serum antibody levels were significantly increased at Day 14 and continued until 42 days post-vaccination. The mRNA expression levels of immune-related genes (IFN-1, TLR22, IL-1β, MHC I, Mx and IgM) were significantly upregulated in liver, spleen, kidney and hindgut after immunization. Four weeks post-immunization, fish were challenged with virulent GCRV by intraperitoneal injection. The results of this challenge study show that orally immunized group exhibited a survival rate of 60% and relative percent survival (RPS) of 56%, whereas the control group had a survival rate of 13% and RPS of 4%. Taken together, our results demonstrate that the silkworm pupae powder containing baculovirus-expressed VP35-VP4 proteins could induce both non-specific and specific immune responses and protect grass carp against GCRV infection, suggesting it could be used as an oral vaccine.

Rabies is a devastating disease affecting almost all mammalian animal species including humans. Vaccines are available to combat the disease. Protection against the disease is rendered by assessing the humoral immune response. Recent reports suggest the role of cell mediated immune response (CMI) in assessing vaccine efficacy. In the present study, two live vectored vaccine candidates containing glycoprotein G of rabies virus were generated using the mesogenic Newcastle disease virus (NDV) strain R2B and another with NDV with an altered fusion protein cleavage site as backbones. The efficacy of these vaccine candidates on testing in experimental mouse model indicated generation of robust humoral and CMI responses. The recombinant NDV containing the altered fusion protein cleavage site with glycoprotein G showed the highest CMI response in mice indicating its usage as a potential live vectored vaccine candidate against the disease., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The grass carp hemorrhagic disease, caused by the grass carp reovirus (GCRV), has resulted in severe economic losses in the aquaculture industry in China. VP4 and VP35 are outer capsid proteins of GCRV and can induce an immune response in the host. Here, three recombinant baculoviruses, AcMNPV-VP35, AcMNPV-VP4, and AcMNPV-VP35-VP4, were generated to express recombinant VP4 and VP35 proteins from GCRV type II in insect cells by using the Bac-to-Bac baculovirus expression system to create a novel subunit vaccine. The expression of recombinant VP35, VP4, and VP35-VP4 proteins in Sf-9 cells were confirmed by Western blotting and immunofluorescence. Recombinant VP35, VP4, and VP35-VP4 were purified from baculovirus-infected cell lysates and injected intraperitoneally (3 μg/fish) into the model rare minnow, Gobiocypris rarus . After 21 days, the immunized fish were challenged with virulent GCRV. Liver, spleen, and kidney samples were collected at different time intervals to evaluate the protective efficacy of the subunit vaccines. The mRNA expression levels of some immune-related genes detected by using quantitative real-time PCR (qRT-PCR) were significantly upregulated in the liver, spleen, and kidney, with higher expression levels in the VP35-VP4 group. The nonvaccinated fish group showed 100% mortality, whereas the VP35-VP4, VP4, and VP35 groups exhibited 67%, 60%, and 33% survival, respectively. In conclusion, our results revealed that recombinant VP35 and VP4 can induce immunity and protect against GCRV infection, with their combined use providing the best effect. Therefore, VP35 and VP4 proteins can be used as a novel subunit vaccine against GCRV infection.

Introduction: Substantial healthcare resources have been diverted to manage the effects of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, and nonemergency neurosurgery has been effectively closed. As we begin to emerge from the crisis, we will need to manage the backlog of nonemergency neurosurgical patients whose treatment has been delayed and remain responsive to further possible surges of SARS-CoV-2 infections., Methods: In the present study, we aimed to identify the core themes and challenges that will limit resumption of a normal neurosurgical service after the SARS-CoV-2 pandemic and to provide pragmatic advice and solutions that could be of utility to clinicians seeking to resume nonemergency neurosurgical care. We reviewed the relevant international policies, a wide range of journalistic and media sources, and expert opinion documents to address the stated aims., Results: We have presented and discussed a range of factors that could become potential barriers to resuming full elective neurosurgical provision and important steps that must be completed to achieve pre-SARS-CoV-2 surgical capacity. We also explored how these challenges can be overcome and outlined the key requirements for a successful neurosurgical exit strategy from the pandemic., Conclusion: The performance of nonemergency neurosurgery can start once minimum criteria have been fulfilled: 1) a structured prioritization of surgical cases; 2) virus infection incidence decreased sufficiently to release previously diverted healthcare resources; 3) adequate safety criteria met for patients and staff, including sufficient personal protective equipment and robust testing availability; and 4) maintenance of systems for rapid communication at organizational and individual levels., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objective: Stereoelectroencephalography (SEEG) is a procedure in which many electrodes are stereotactically implanted within different regions of the brain to estimate the epileptogenic zone in patients with drug-refractory focal epilepsy. Computer-assisted planning (CAP) improves risk scores, gray matter sampling, orthogonal drilling angles to the skull and intracerebral length in a fraction of the time required for manual planning. Due to differences in planning practices, such algorithms may not be generalizable between institutions. We provide a prospective validation of clinically feasible trajectories using "spatial priors" derived from previous implantations and implement a machine learning classifier to adapt to evolving planning practices. Methods: Thirty-two patients underwent consecutive SEEG implantations utilizing computer-assisted planning over 2 years. Implanted electrodes from the first 12 patients (108 electrodes) were used as a training set from which entry and target point spatial priors were generated. CAP was then prospectively performed using the spatial priors in a further test set of 20 patients (210 electrodes). A K-nearest neighbor (K-NN) machine learning classifier was implemented as an adaptive learning method to modify the spatial priors dynamically. Results: All of the 318 prospective computer-assisted planned electrodes were implanted without complication. Spatial priors developed from the training set generated clinically feasible trajectories in 79% of the test set. The remaining 21% required entry or target points outside of the spatial priors. The K-NN classifier was able to dynamically model real-time changes in the spatial priors in order to adapt to the evolving planning requirements. Conclusions: We provide spatial priors for common SEEG trajectories that prospectively integrate clinically feasible trajectory planning practices from previous SEEG implantations. This allows institutional SEEG experience to be incorporated and used to guide future implantations. The deployment of a K-NN classifier may improve the generalisability of the algorithm by dynamically modifying the spatial priors in real-time as further implantations are performed., (Copyright © 2020 Vakharia, Sparks, Granados, Miserocchi, McEvoy, Ourselin and Duncan.)

The past decade has seen a significant shift in the number of centers performing intracranial electroencephalography from subdural grids and strips to stereoelectroencephalography (SEEG). Unlike grid and strip insertion or other stereotactic procedures in which the cortical surface is visualized, SEEG involves insertion of an electrode through a bolt anchored into the skull. Due to the multidisciplinary nature of SEEG trajectory planning, it often is time-consuming and iterative. Computer-assisted planning improves time taken and efficacy of SEEG trajectory planning. This article provides an overview of the considerations, controversies, and practicalities of implementing an automated computer-assisted planning solution for SEEG planning., Competing Interests: Disclosure The authors have nothing to disclose., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Viral hemorrhagic septicemia virus (VHSV) is one of the most deadly infectious fish pathogens, posing a serious threat to the aquaculture industry and freshwater ecosystems worldwide. Previous work showed that VHSV sub-genotype IVb suppresses host innate immune responses, but the exact mechanism by which VHSV IVb inhibits antiviral response remains incompletely characterized. As with other novirhabdoviruses, VHSV IVb contains a unique and highly variable nonvirion (NV) gene, which is implicated in viral replication, virus-induced apoptosis and regulating interferon (IFN) production. However, the molecular mechanisms underlying the role of IVb NV gene in regulating viral or cellular processes is poorly understood. Compared to the wild-type recombinant (rWT) VHSV, mutant VHSV lacking a functional IVb NV reduced IFN expression and compromised innate immune response of the host cells by inhibiting translation. VHSV IVb infection increased phosphorylated eukaryotic initiation factor 2α (p-eIF2α), resulting in host translation shutoff. However, VHSV IVb protein synthesis proceeds despite increasing phosphorylation of eIF2α. During VHSV IVb infection, eIF2α phosphorylation was mediated via PKR-like endoplasmic reticulum kinase (PERK) and was required for efficient viral protein synthesis, but shutoff of host translation and IFN signaling was independent of p-eIF2α. Similarly, IVb NV null VHSV infection induced less p-eIF2α, but exhibited decreased viral protein synthesis despite increased levels of viral mRNA. These findings show a role for IVb NV in VHSV pathogenesis by utilizing the PERK-eIF2α pathway for viral-mediated host shutoff and interferon signaling to regulate host cell response.

Background: Stereotactic brain biopsy is among the most common neurosurgical procedures. Planning an optimally safe surgical trajectory requires careful attention to a number of features including the following: (1) traversing the skull perpendicularly; (2) minimizing trajectory length; and (3) avoiding critical neurovascular structures., Objective: To evaluate a platform, SurgiNav, for automated trajectory planning in stereotactic brain biopsy., Methods: A prospectively maintained database was searched between February and August 2017 to identify all adult patients who underwent stereotactic brain biopsy and for whom postoperative imaging was available. In each case, the standard preoperative, T1-weighted, gadolinium-enhanced magnetic resonance imaging was used to generate a model of the cortex. A surgical trajectory was then generated using computer-assisted planning (CAP) , and metrics of the trajectory were compared to the trajectory of the previously implemented manual plan (MP)., Results: Fifteen consecutive patients were identified. Feasible trajectories were generated using CAP in all patients, and the mean angle determined using CAP was more perpendicular to the skull than using MP (10.0° vs 14.6° from orthogonal; P = .07), the mean trajectory length was shorter (38.5 vs 43.5 mm; P = .01), and the risk score was lower (0.27 vs 0.52; P = .03)., Conclusion: CAP for stereotactic brain biopsy appears feasible and may be safer in selected cases., (© Congress of Neurological Surgeons 2019.)

Background: Anterior two-thirds corpus callosotomy is an effective palliative neurosurgical procedure for drug-refractory epilepsy that is most commonly used to treat drop-attacks. Laser interstitial thermal therapy is a novel stereotactic ablative technique that has been utilised as a minimally invasive alternative to resective and disconnective open neurosurgery. Case series have reported success in performing laser anterior two-thirds corpus callosotomy. Computer-assisted planning algorithms may help to automate and optimise multi-trajectory planning for this procedure., Objective: To undertake a simulation-based feasibility study of computer-assisted corpus callostomy planning in comparison with expert manual plans in the same patients., Methods: Ten patients were selected from a prospectively maintained database. Patients had previously undergone diffusion-weighted imaging and digital subtraction angiography as part of routine SEEG care. Computer-assisted planning was performed using the EpiNav™ platform and compared to manually planned trajectories from two independent blinded experts. Estimated ablation cavities were used in conjunction with probabilistic tractography to simulate the expected extent of interhemispheric disconnection., Results: Computer-assisted planning resulted in significantly improved trajectory safety metrics (risk score and minimum distance to vasculature) compared to blinded external expert manual plans. Probabilistic tractography revealed residual interhemispheric connectivity in 1/10 cases following computer-assisted planning compared to 4/10 and 2/10 cases with manual planning., Conclusion: Computer-assisted planning successfully generates multi-trajectory plans capable of LITT anterior two-thirds corpus callosotomy. Computer-assisted planning may provide a means of standardising trajectory planning and serves as a potential new tool for optimising trajectories. A prospective validation study is now required to determine if this translates into improved patient outcomes., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Although typical Newcastle disease virus (NDV) vaccines can prevent mortality, they are not effective in preventing viral shedding. To overcome this, genotype-matched vaccines have been proposed. To date, this approach has never been tested against genotype XII strains. In this study, we generated and assessed the protection against genotype XII challenge of two chimeric NDV vaccine strains (rLS1-XII-1 and rLS1-XII-2). The rLS1-XII-1 virus has the complete fusion protein (F) and the hemagglutinin-neuraminidase (HN) open reading frames replaced with those from genotype XII strain NDV/peacock/Peru/2011 (PP2011) in a recombinant LaSota (rLS1) backbone. In rLS1-XII-2 virus, cytoplasmic tails of F and HN proteins were restored to those of rLS1. In vitro evaluation showed that rLS1-XII-2 and the parental rLS1 strains replicate at higher efficiencies than rLS1-XII-1. In the first vaccine/challenge experiment, SPF chickens vaccinated with rLS1-XII-1 virus showed only 71.3% protection, whereas, rLS1 and rLS1-XII-2 vaccinated chickens were fully protected. In a second experiment, both rLS1-XII-2 and the commercial vaccine strain LaSota induced 100% protection. However, rLS1-XII-2 virus significantly reduced viral shedding, both in the number of shedding birds and in quantity of shed virus. In conclusion, we have developed a vaccine candidate capable of fully protecting chickens against genotype XII challenges. Furthermore, we have shown the importance of cytoplasmic tails in virus replication and vaccine competence., Competing Interests: MF-D is the Founder and CEO of FARVET S.A.C. All other authors declare having no 494 competing interests (RI-L, AC, KC and VNV). This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Stereoelectroencephalography (SEEG) is a diagnostic procedure in which multiple electrodes are stereotactically implanted within predefined areas of the brain to identify the seizure onset zone, which needs to be removed to achieve remission of focal epilepsy. Computer-assisted planning (CAP) has been shown to improve trajectory safety metrics and generate clinically feasible trajectories in a fraction of the time needed for manual planning. We report a prospective validation study of the use of EpiNav (UCL, London, UK) as a clinical decision support software for SEEG. Thirteen consecutive patients (125 electrodes) undergoing SEEG were prospectively recruited. EpiNav was used to generate 3D models of critical structures (including vasculature) and other important regions of interest. Manual planning utilizing the same 3D models was performed in advance of CAP. CAP was subsequently employed to automatically generate a plan for each patient. The treating neurosurgeon was able to modify CAP generated plans based on their preference. The plan with the lowest risk score metric was stereotactically implanted. In all cases (13/13), the final CAP generated plan returned a lower mean risk score and was stereotactically implanted. No complication or adverse event occurred. CAP trajectories were generated in 30% of the time with significantly lower risk scores compared to manually generated. EpiNav has successfully been integrated as a clinical decision support software (CDSS) into the clinical pathway for SEEG implantations at our institution. To our knowledge, this is the first prospective study of a complex CDSS in stereotactic neurosurgery and provides the highest level of evidence to date.

Newcastle disease (ND) and avian reovirus (ARV) infections are a serious threat to the poultry industry, which causes heavy economic losses. The mesogenic NDV strain R2B is commonly used as a booster vaccine in many Asian countries to control the disease. In this seminal work, a recombinant NDV strain R2B expressing the sigma C (σC) gene of ARV (rNDV-R2B-σC) was generated by reverse genetics, characterized in vitro and tested as a bivalent vaccine candidate in chickens. The recombinant rNDV-R2B-σC virus was attenuated as compared to the parent rNDV-R2B virus as revealed by standard pathogenicity assays. The generated vaccine candidate, rNDV-R2B-σC, could induce both humoral and cell mediated immune responses in birds and gave complete protection against virulent NDV and ARV challenges. Post-challenge virus shedding analysis revealed a drastic reduction in NDV shed, as compared to unvaccinated birds.

Objective: Various forms of vascular imaging are performed to identify vessels that should be avoided during stereoelectroencephalography (SEEG) planning. Digital subtraction angiography (DSA) is the gold standard for intracranial vascular imaging. DSA is an invasive investigation, and a balance is necessary to identify all clinically relevant vessels and not to visualize irrelevant vessels that may unnecessarily restrict electrode placement. We sought to estimate the size of vessels that are clinically significant for SEEG planning., Methods: Thirty-three consecutive patients who underwent 354 SEEG electrode implantations planned with computer-assisted planning and DSA segmentation between 2016 and 2018 were identified from a prospectively maintained database. Intracranial positions of electrodes were segmented from postimplantation computed tomography scans. Each electrode was manually reviewed using "probe-eye view" with the raw preoperative DSA images for vascular conflicts. The diameter of vessels and the location of conflicts were noted. Vessel conflicts identified on raw DSA images were cross-referenced against other modalities to determine whether the conflict could have been detected., Results: One hundred sixty-six vessel conflicts were identified between electrodes and DSA-identified vessels, with 0-3 conflicts per electrode and a median of four conflicts per patient. The median diameter of conflicting vessels was 1.3 mm (interquartile range [IQR] = 1.0-1.5 mm). The median depth of conflict was 31.0 mm (IQR = 14.3-45.0 mm) from the cortical surface. The addition of sulcal models to DSA, magnetic resonance venography (MRV), and T1 + gadolinium images, as an exclusion zone during computer-assisted planning, would have prevented the majority of vessel conflicts. We were unable to determine whether vessels were displaced or transected by the electrodes., Significance: Vascular segmentation from DSA images was significantly more sensitive than T1 + gadolinium or MRV images. Electrode conflicts with vessels 1-1.5 mm in size did not result in a radiologically detectable or clinically significant hemorrhage and could potentially be excluded from consideration during SEEG planning., (© 2019 The Authors. Epilepsia published by Wiley Periodicals, Inc. on behalf of International League Against Epilepsy.)

Objective: Laser interstitial thermal therapy (LITT) is a novel minimally invasive alternative to open mesial temporal resection in drug-resistant mesial temporal lobe epilepsy (MTLE). The safety and efficacy of the procedure are dependent on the preplanned trajectory and the extent of the planned ablation achieved. Ablation of the mesial hippocampal head has been suggested to be an independent predictor of seizure freedom, whereas sparing of collateral structures is thought to result in improved neuropsychological outcomes. We aim to validate an automated trajectory planning platform against manually planned trajectories to objectively standardize the process., Methods: Using the EpiNav platform, we compare automated trajectory planning parameters derived from expert opinion and machine learning to undertake a multicenter validation against manually planned and implemented trajectories in 95 patients with MTLE. We estimate ablation volumes of regions of interest and quantify the size of the avascular corridor through the use of a risk score as a marker of safety. We also undertake blinded external expert feasibility and preference ratings., Results: Automated trajectory planning employs complex algorithms to maximize ablation of the mesial hippocampal head and amygdala, while sparing the parahippocampal gyrus. Automated trajectories resulted in significantly lower calculated risk scores and greater amygdala ablation percentage, whereas overall hippocampal ablation percentage did not differ significantly. In addition, estimated damage to collateral structures was reduced. Blinded external expert raters were significantly more likely to prefer automated to manually planned trajectories., Significance: Retrospective studies of automated trajectory planning show much promise in improving safety parameters and ablation volumes during LITT for MTLE. Multicenter validation provides evidence that the algorithm is robust, and blinded external expert ratings indicate that the trajectories are clinically feasible. Prospective validation studies are now required to determine if automated trajectories translate into improved seizure freedom rates and reduced neuropsychological deficits., (© 2019 The Authors. Epilepsia published by Wiley Periodicals, Inc. on behalf of International League Against Epilepsy.)

Viral hemorrhagic septicemia virus (VHSV), a fish rhabdovirus, infects several marine and freshwater fish species. There are many strains of VHSV that affect different fish, but some strains of one genetic subgroup have gained high virulence in rainbow trout ( Oncorhynchus mykiss ). To define the genetic basis of high virulence in trout, we used reverse genetics to create chimeric VHSVs in which viral nucleoprotein (N), P (phosphoprotein), or M (matrix protein) genes, or the N and P genes, were exchanged between a trout-virulent European VHSV strain (DK-3592B) and a trout-avirulent North American VHSV strain (MI03). Testing of the chimeric recombinant VHSV (rVHSV) by intraperitoneal injection in juvenile rainbow trout showed that exchanges of the viral P or M genes had no effect on the trout virulence phenotype of either parental strain. However, reciprocal exchanges of the viral N gene resulted in a partial gain of function in the chimeric trout-avirulent strain (22% mortality) and complete loss of virulence for the chimeric trout-virulent strain (2% mortality). Reciprocal exchanges of both the N and P genes together resulted in complete gain of function in the chimeric avirulent strain (82% mortality), again with complete loss of virulence in the chimeric trout-virulent strain (0% mortality). Thus, the VHSV N gene contains an essential determinant of trout virulence that is strongly enhanced by the viral P gene. We hypothesize that the host-specific virulence mechanism may involve increased efficiency of the viral polymerase complex when the N and P proteins have adapted to more efficient interaction with a host component from rainbow trout. IMPORTANCE Rainbow trout farming is a major food source industry worldwide that has suffered great economic losses due to host jumps of fish rhabdovirus pathogens, followed by evolution of dramatic increases in trout-specific virulence. However, the genetic determinants of host jumps and increased virulence in rainbow trout are unknown for any fish rhabdovirus. Previous attempts to identify the viral genes containing trout virulence determinants of viral hemorrhagic septicemia virus (VHSV) have not been successful. We show here that, somewhat surprisingly, the viral nucleocapsid (N) and phosphoprotein (P) genes together contain the determinants responsible for trout virulence in VHSV. This suggests a novel host-specific virulence mechanism involving the viral polymerase and a host component. This differs from the known virulence mechanisms of mammalian rhabdoviruses based on the viral P or M (matrix) protein., (Copyright © 2019 American Society for Microbiology.)

Background: Stereotactic neurosurgical procedures carry a risk of intracranial hemorrhage, which may result in significant morbidity and mortality. Vascular imaging is crucial for planning stereotactic procedures to prevent conflicts with intracranial vasculature. There is a wide range of vascular imaging methods used for stereoelectroencephalography (SEEG) trajectory planning. Computer-assisted planning (CAP) improves planning time and trajectory metrics. We aimed to quantify the effect of different vascular imaging protocols on CAP trajectories for SEEG., Methods: Ten patients who had undergone SEEG (95 electrodes) following preoperative acquisition of gadolinium-enhanced magnetic resonance imaging (MR + Gad), magnetic resonance angiography and magnetic resonance angiography (MRV + MRA), and digital subtraction catheter angiography (DSA) were identified from a prospectively maintained database. SEEG implantations were planned using CAP using DSA segmentations as the gold standard. Strategies were then recreated using MRV + MRA and MR + Gad to define the "apparent" and "true" risk scores associated with each modality. Vessels of varying diameter were then iteratively removed from the DSA segmentation to identify the size at which all 3 vascular modalities returned the same safety metrics., Results: CAP performed using DSA vessel segmentations resulted in significantly lower "true" risk scores and greater minimum distances from vasculature compared with the "true" risk associated with MR + Gad and MRV + MRA. MRV + MRA and MR + Gad returned similar risk scores to DSA when vessels <2 mm and <4 mm were not considered, respectively., Conclusions: Significant variability in vascular imaging and trajectory planning practices exist for SEEG. CAP performed with MR + Gad or MRV + MRA alone returns "falsely" lower risk scores compared with DSA. It is unclear whether DSA is oversensitive and thus restricting potential trajectories., (© 2019 The Authors.)

Objective: The accuracy of stereoelectroencephalography (SEEG) electrode implantation is an important factor in maximizing its safety. The authors established a quality assurance (QA) process to aid advances in implantation accuracy., Methods: The accuracy of three consecutive modifications of a frameless implantation technique was quantified in three cohorts comprising 22, 8, and 23 consecutive patients. The modifications of the technique aimed to increase accuracy of the bolt placement., Results: The lateral shift of the axis of the implanted bolt at the level of the planned entry point was reduced from a mean of 3.0 ± 1.6 mm to 1.4 ± 0.8 mm. The lateral shift of the axis of the implanted bolt at the level of the planned target point was reduced from a mean of 3.8 ± 2.5 mm to 1.6 ± 0.9 mm., Conclusions: This QA framework helped to isolate and quantify the factors introducing inaccuracy in SEEG implantation, and to monitor ongoing accuracy and the effect of technique modifications.

Background: Viral hemorrhagic septicemia virus (VHSV), a fish rhabdovirus belonging to the Novirhabdovirus genus, causes severe disease and mortality in many marine and freshwater fish species worldwide. VHSV isolates are classified into four genotypes and each group is endemic to specific geographic regions in the north Atlantic and Pacific Oceans. Most viruses in the European VHSV genotype Ia are highly virulent for rainbow trout (Oncorhynchus mykiss), whereas, VHSV genotype IVb viruses from the Great Lakes region in the United States, which caused high mortality in wild freshwater fish species, are avirulent for trout. This study describes molecular characterization and construction of an infectious clone of the virulent VHSV-Ia strain DK-3592B from Denmark, and application of the clone in reverse genetics to investigate the role of selected VHSV protein(s) in host-specific virulence in rainbow trout (referred to as trout-virulence)., Methods: Overlapping cDNA fragments of the DK-3592B genome were cloned after RT-PCR amplification, and their DNA sequenced by the di-deoxy chain termination method. A full-length cDNA copy (pVHSVdk) of the DK-3592B strain genome was constructed by assembling six overlapping cDNA fragments by using natural or artificially created unique restriction sites in the overlapping regions of the clones. Using an existing clone of the trout-avirulent VHSV-IVb strain MI03 (pVHSVmi), eight chimeric VHSV clones were constructed in which the coding region(s) of the glycoprotein (G), non-virion protein (NV), G and NV, or G, NV and L (polymerase) genes together, were exchanged between the two clones. Ten recombinant VHSVs (rVHSVs) were generated, including two parental rVHSVs, by transfecting fish cells with ten individual full-length plasmid constructs along with supporting plasmids using the established protocol. Recovered rVHSVs were characterized for viability and growth in vitro and used to challenge groups of juvenile rainbow trout by intraperitoneal injection., Results: Complete sequence of the VHSV DK-3592B genome was determined from the cloned cDNA and deposited in GenBank under the accession no. KC778774. The trout-virulent DK-3592B genome (genotype Ia) is 11,159 nt in length and differs from the trout-avirulent MI03 genome (pVHSVmi) by 13% at the nucleotide level. When the rVHSVs were assessed for the trout-virulence phenotype in vivo, the parental rVHSVdk and rVHSVmi were virulent and avirulent, respectively, as expected. Four chimeric rVHSVdk viruses with the substitutions of the G, NV, G and NV, or G, NV and L genes from the avirulent pVHSVmi constructs were still highly virulent (100% mortality), while the reciprocal four chimeric rVHSVmi viruses with genes from pVHSVdk remained avirulent (0-10% mortality)., Conclusions: When chimeric rVHSVs, containing all the G, NV, and L gene substitutions, were tested in vivo, they did not exhibit any change in trout-virulence relative to the background clones. These results demonstrate that the G, NV and L genes of VHSV are not, by themselves or in combination, major determinants of host-specific virulence in trout.

White Tail Disease (WTD) is one of the important viral diseases of fresh water giant prawn Macrobrachium rosenbergii, which is caused by Macrobrachium rosenbergii nodavirus (MrNV). In the present study, the capsid protein gene of MrNV containing a His-tag was cloned into a baculovirus vector pVL1393 and expressed the recombinant MrNV protein in insect cells, using a baculovirus expression system. A band corresponding to the MrNV protein of 43 kDa was characterized after fractionating the proteins of baculovirus-infected cell lysates by SDS-polyacrylamide gel, and immunostaining with His-tag monoclonal antibody. Furthermore, purified MrNV capsid protein assembled into virus-like particles (VLPs) of ∼30 nm in diameter, when examined by transmission electron microscopy (TEM). To vaccinate the larvae by oral route, the recombinant MrNV (r-MrNV) protein was coated with artificial prawn feed and fed to M. rosenbergii larvae (90 ± 10 mg) for 60 days. After 30 and 60 days of vaccine treatment, group of prawns were challenged with virulent MrNV orally. Samples were collected at different time intervals to evaluate the survival of larvae and to analyze the presence of MrNV by double-step PCR and expression of immune/ toll-like receptor (TLR) genes. Non-vaccinated group of M. rosenbergii larvae succumbed to death and had 90% mortality, whereas the r-MrNV protein treated groups exhibited 65 and 80% survival (P  ≤  0.001) for 30 and 60 days post-vaccination (dpv), respectively. Double-step PCR diagnosis revealed that there was 100% positive signals observed in non-vaccinated prawn group, whereas the infection was reduced significantly (P < 0.001) to 32 and 17% respectively in 30 and 60 dpv. Among the four different immune/ TLR genes such as antimicrobial peptide (Mramp), lysozyme (MrLY), proPhenol Oxidase (MrPPO) and Toll-Like Receptor (MrToll) expression screening, Mramp was successfully expressed in the MrNV subunit protein vaccinated prawns, whereas the non-vaccinated prawn had no immune/TLR gene expression. Taken together, our results demonstrate that oral vaccination of M. rosenbergii larvae with baculovirus-expressed MrNV capsid protein confer up to 78% protection against MrNV infection., (Copyright © 2018 Elsevier Ltd. All rights reserved.)