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Cobweb disease caused by members of the Cladobotryum genus is a major problem for growers of the white button mushroom (Agaricus bisporus). Synthetic fungicides such as prochloraz and metrafenone have been very successful at targeting and eliminating the pathogens that cause mushroom disease. However, prochloraz can no longer be used in the European Union (EU) from June 2023 and over-reliance on metrafenone has resulted in putative resistant pathogenic strains emerging. Prochloraz still showed good control of two different isolates of Cladobotryum mycophilum with efficacy values consistently reaching 70%. Metrafenone inhibited the growth of C. mycophilum isolate 618, which was isolated before metrafenone was introduced (efficacy 96%), but it failed to control C. mycophilum 1546, which was isolated after metrafenone was introduced, and which should now be classified as resistant. Two further C. mycophilum isolates from mushroom farms in 2019 also showed metrafenone resistance in vitro. In this work two biological control agents (BCAs) were investigated as potential environmentally sustainable alternatives to the fungicides prochloraz and metrafenone. The BCA Bacillus velezensis QST 713 was unsuccessful in controlling cobweb disease caused by C. mycophilum isolate 1546 while the BCA Bacillus velezensis Kos showed moderate control over two trials reaching 30–40% efficacy. Lower inoculum concentrations resulted in slightly lower but not significantly different disease levels across all treatments. Future trials with BCAs need to look at alternative methods to evaluate efficacy. • Metrafenone - resistant Cobweb isolates were isolated from mushroom crops. • Bacillus velezensis QST 713 was unsuccessful in controlling cobweb disease. • Bacillus velezensis Kos culture filtrate had limited effect on cobweb disease. • Reducing inoculum concentration by 50% did not significantly reduce disease levels. [ABSTRACT FROM AUTHOR]

• Functional properties of excitatory inputs onto cortical neurons are more diverse. • Cortical neurons can receive imbalanced synaptic excitation and inhibition. • Current theoretical models are consistent with these findings. • New evidence suggests cortical circuits support operational flexibility. Establishing the fundamental principles that underlie the integration of excitatory and inhibitory presynaptic input populations is crucial to understanding how individual cortical neurons transform signals from peripheral receptors. Here we review recent studies using novel tools to examine the functional properties of excitatory synaptic inputs and the tuning of excitation and inhibition onto individual neurons. New evidence challenges existing synaptic connectivity rules and suggests a more complex functional synaptic architecture that supports a broad range of operations, enabling single neurons to encode multiple sensory features and flexibly shape their computations in the face of diverse sensory input. [ABSTRACT FROM AUTHOR]

Impaction bone grafting is a method of restoring bone stock to patients who have suffered significant bone loss due to revision total hip surgery. The procedure requires morsellised cancellous bone (MCB) to be impacted into the site of bone loss in order to stabilise the prosthesis with the aim of long term resorption and reintegration of the impacted bone graft. Due to financial cost and the potential to transmit disease, the use of supplementary material, known as an extender, is frequently used to increase the graft material volume. This study investigates the use of hardened Hydroset (Stryker Corp, MA, USA), an injectable bone cement (IBC), as an extender material and compares the performance of the IBC in different weight percent inclusions to a commercially available bone graft extender (GCP, BoneSave, Stryker Corp, MA, USA). The surgical impaction procedure was standardised and samples were evaluated in terms of graft stiffness and height. It was observed that 30 wt% IBC extended samples had significantly improved graft stiffness ( p = 0.02) and no significant different in height ( p = 0.067) over a 100% MCB control sample. Cyclic loading, representative of gait, found that the IBC subsided similarly to the commercial bone substitute in wt% above 10%. Shear testing of the impacted grafts showed no significant differences between GCP and IBC with impaction forces determining the shear parameters of impacted grafts. The effects of the impaction and cyclical loading procedures on extender particle sizes was assessed via particle size analysis. It was found that the IBC extended samples demonstrated reduced friability, evident in the better retention of particle size as a result of both impaction and gait representative loading compared to that of the GCP samples. This indicates a potential reduction in issues arising from small particle migration to joint surfaces. Scanning electron microscopy of the MCB particles with both GCP and IBC as extenders showed retention of the porous trabecular structure post-testing which is essential for revascularisation and bone growth into the graft. [ABSTRACT FROM AUTHOR]

Impaction bone grafting is a method of restoring bone stock to patients suffering significant bone loss due to revision total hip surgery. The procedure requires morselised bone (MB) to be impacted into the site of bone loss in order to stabilise the prosthesis with the aim of the long term resorption and reintegration of the impacted bone graft. Currently, the method for producing MB requires the use of expensive surgical bone mills or manually-intensive rongeurs that can produce a limited variety of particle sizes and may have a low throughput. This study examines the potential to produce suitable MB using a domestic blender. The method produces a wide range of particle sizes without the need for an adjustment of the system. It was found through packing modelling that this particle distribution resulted in reduced initial graft porosity and thus a theoretical potential to increase the graft stiffness and ability of the graft to stabilise a prosthesis in comparison to a manually prepared roughly cut morselised bone samples. Mechanical testing confirmed the increased mechanical performance of the graft through both impaction testing and subsidence testing. The blended MB was found to exhibit greater graft stiffness under the same impaction conditions. The graft was also found to have subsided less in comparison to the rough cut, less well graded MB. Scanning electron imaging also confirmed the retention of the trabecular structure necessary for revascularisation and host bone ingrowth. In conclusion, the blender method offers a rapid and cheap way of obtaining morselised bone with favourable particle size distribution, particle morphology and mechanical properties with preservation of the bone trabecular structure. [ABSTRACT FROM AUTHOR]

Atypical meningiomas have a greater propensity to recur than benign meningiomas and the benefits of early adjuvant radiotherapy are unclear. Existing studies report conflicting results. This retrospective cohort study evaluated the role of early adjuvant radiotherapy following surgical resection of atypical meningioma. A triple center case-note review of adults with newly-diagnosed atypical meningiomas between 2001 and 2010 was performed. Pathology diagnosis was made according to the World Health Organization classification in use at the time of surgery. Patients with multiple meningiomas, neurofibromatosis type 2 and radiation-induced meningiomas were excluded. Extent of resection was defined as gross total resection (GTR; Simpson Grade I–III) or subtotal resection (STR; Simpson Grade IV–V). Survival analysis was performed using the Kaplan–Meier method. One hundred thirty-three patients were identified with a median age of 62 years (range 22–86 years) and median follow-up of 57.4 months (range 0.1–152.2 months). Tumors were mostly located in the convexity (50.4%) or falcine/parasagittal regions (27.1%). GTR (achieved in 85%) was associated with longer progression free survival (PFS) (5 year PFS 81.2% versus 40.08%, log-rank = 11.117, p = 0.001) but not overall survival (OS) (5 year OS 76.6% versus 39.7%, log-rank = 3.652, p = 0.056). Following GTR, early adjuvant radiotherapy was administered to 28.3% of patients and did not influence OS (5 year OS 77.0% versus 75.7%, log-rank = 0.075, p = 0.784) or PFS (5 year PFS 82.0% versus 79.3%, log-rank = 0.059, p = 0.808). Although extent of resection emerged as an important prognostic variable, early adjuvant radiotherapy did not influence outcome following GTR of atypical meningiomas. Prospective randomized controlled trials are planned. [ABSTRACT FROM AUTHOR]

Microphthalmia, anophthalmia and coloboma (MAC) are distinct phenotypes that represent a continuum of structural developmental eye defects. In severe bilateral cases (anophthalmia or severe microphthalmia) the genetic cause is now identifiable in approximately 80 percent of cases, with de novo heterozygous loss-of-function mutations in SOX2 or OTX2 being the most common. The genetic cause of other forms of MAC, in particular isolated coloboma, remains unknown in the majority of cases. This review will focus on MAC phenotypes that are associated with mutation of the genes SOX2, OTX2, PAX6, STRA6, ALDH1A3, RARB, VSX2, RAX, FOXE3, BMP4, BMP7, GDF3, GDF6, ABCB6, ATOH7, C12orf57, TENM3 (ODZ3), and VAX1. Recently reported mutation of the SALL2 and YAP1 genes are discussed in brief. Clinical and genetic features were reviewed in a total of 283 unrelated MAC cases or families that were mutation-positive from these 20 genes. Both the relative frequency of mutations in MAC cohort screens and the level of confidence in the assignment of disease-causing status were evaluated for each gene. [ABSTRACT FROM AUTHOR]

Anaphylaxis is a challenging condition for any austere environment. It is unpredictable, has sudden onset and a high fatality rate, and is responsive only to epinephrine, a prescription medication. The Wilderness Medical Society has formally recommended that non-medical providers working in austere environments be trained to administer epinephrine. Medical providers frequently prescribe auto-injectors for this purpose due to their ease of use by nonmedical providers. However, auto-injectors have limitations in the wilderness environment, particularly due to their single-dose (or at most 2-dose) design. This paper describes an austere environment technique for obtaining multiple additional doses of epinephrine from auto-injectors that have already been used as designed. [ABSTRACT FROM AUTHOR]

Background: Analysis of naturally occurring T regulatory CD4+ (nTreg) cells in human diseases is hampered by the lack of specific surface marker. Indeed, the CD25 antigen, which is typically used to identify nTreg cells, is also expressed on activated effector T cells. Objective: We sought to examine whether CD4+ T cells bearing CD103 are suppressor cells, regardless of CD25 coexpression. Methods: We first compared freshly isolated tonsillar CD103+CD25− cells with their CD103−CD25high counterparts for their capacity to suppress T-cell response and their expression of FoxP3 mRNA. Next CD103 was induced on neonatal or adult CD4+ T cells stimulated with allogeneic dendritic cells, and the CD103+ and CD103− fractions were compared as above. Results: Tonsillar CD4+CD103+CD25− T cells displayed comparable suppressive activity and contained similar amounts of FoxP3 mRNA as their CD103−CD25high counterparts. In vitro–generated alloantigen-primed CD103+ cells coexpressed CD25, suppressed T-cell activation, and contained more FoxP3 mRNA than the CD103−CD25+ cells isolated from the same cultures. Finally, neonatal alloreactive cells contained more CD103+ Treg cells than their adult counterparts and, unlike the latter, became hyporesponsive to the priming alloantigens. Conclusions: The examination of CD103 and CD25 coexpression allows identification of 3 subsets of human CD4+ nTreg cells, and the detection of CD103 on CD4+ T cells identifies nTreg cells, regardless of CD25 coexpression. Clinical implications: The greater induction of CD103+ suppressor cells by cord blood should be related to its successful clinical use as an alternative to adult bone marrow transplantation. [Copyright &y& Elsevier]

In developed countries, malformations of the eye are among the most common causes of serious visual impairment in newborns. The identification of pathogenic mutations in autosomal and X-linked transcription factors has advanced our understanding of the critical stages in human eye development and has begun to explain some unusual inheritance characteristics of these disorders. The functional characterisation of these genes in model organisms has prompted reinvestigation of affected individuals to identify previously unrecognized but consistent extra-ocular malformations. This dialogue between clinical genetics and basic developmental biology provides a paradigm to enhance our understanding of many critical developmental processes in human embryogenesis. [Copyright &y& Elsevier]

Autosomal trisomy is a common cause of human miscarriage, malformations and learning disability. Primary gene-dosage effects have been confirmed by recent transcriptome analyses. The importance (or existence) of trans-acting effects on disomic genes remains, surprisingly, controversial. In this article, I propose a model of the main genetic mechanisms that are responsible for producing the transcriptional derangement associated with trisomy. This has implications for future study design. [Copyright &y& Elsevier]

Here we review the results of anatomical and physiological studies in tree shrew visual cortex which focus on the contribution of vertical and horizontal inputs to receptive field center and surround properties of layer 2/3 neurons. A fundamental feature of both sets of connections is the arrangement of axon arbors in a fashion that respects both the orientation preference and retinotopic displacement of the target site. As a result, layer 2/3 neurons receive convergent input from populations of layer 4 and other layer 2/3 neurons whose receptive fields are displaced along an axis in visual space that corresponds to their preferred orientation. Although, horizontal connections extend for greater distances across the cortical surface than vertical connections, the majority of these inputs link neurons with overlapping receptive fields, emphasizing that both feed-forward and recurrent circuits are likely to play a constructive role in generating properties (such as orientation selectivity) that define the receptive field center. Both within and beyond the dimensions of the receptive field center, the distribution of horizontal connections accords remarkably well with the magnitude and axial tuning of length summation effects. Taken together, these results suggest a continuum of functional properties that transcends the traditional designation of receptive field center and surround. By extension, we suggest that the perceptual effects of stimulus context may arise from stimulus interactions within the receptive field center as well as between center and surround. [Copyright &y& Elsevier]

Objective. To assess and model the influence of collagen fibre imbalance within the annulus fibrosus on the initiation and progression of adolescent idiopathic scoliosis deformity.Background. A number of aetiological factors have been proposed for idiopathic scoliosis, including a contribution from the intervertebral disc. The specific influence of the annulus fibrosus has yet to be effectively modelled.Methods. A mathematical model was used to determine the contribution of collagen fibre orientation and directional imbalance within the annulus fibrosus to vertebral rotation and overall deformity of thoracic spine. Rotations, due to collagen fibre imbalance, and translations, due to rapid growth, were applied to a simplified model of the thoracic spine, using a three-dimensional transformation matrix approach.Results. The ratio of clockwise to anti-clockwise fibres in the intervertebral disc (from unity to 0.80) influenced the induced rotation. The three-dimensional model illustrates the initiation and progression of the scoliotic deformity during adolescent growth, being most obvious at larger growth rates.Conclusions. Imbalance in the ratio of clockwise and anti-clockwise collagen fibres within the annulus fibrosus has been demonstrated to have the potential to contribute to the progression of scoliosis. For a given fibre ratio, the rate of growth does not influence the induced rotation, but directly influences the severity of the resulting deformity.RelevanceThe model defines the potential contribution of collagen fibre imbalance to adolescent idiopathic scoliosis, supporting the clinical observation that greatest progression of deformity occurs during phases of rapid adolescent growth. However, the underlying mechanism is within the annulus fibrosus, assisting in the search for the responsible genes. [Copyright &y& Elsevier]

Cladobotryum mycophilum , the causative agent of cobweb disease on Agaricus bisporus results in significant crop losses for mushroom growers worldwide. Cobweb disease is treated through strict hygiene control methods and the application of chemical fungicides but an increase in fungicide resistant Cladobotryum strains has resulted in a need to develop alternative biocontrol treatment methods. The aim of the work presented here was to evaluate the response of C. mycophilum to a Bacillus velezensis isolate to assess its potential as a novel biocontrol agent. Exposure of 48 hr C. mycophilum cultures to 25% v/v 96 hr B. velezensis culture filtrate resulted in a 57% reduction in biomass (P < 0.0002), a disruption in hyphal structure and morphology, and the appearance of aurofusarin, a secondary metabolite which is a known indicator of oxidative stress, in culture medium. Proteomic analysis of B. velezensis culture filtrate revealed the presence of peptidase 8 (subtilisin), peptide deformylase and probable cytosol aminopeptidase which are known to induce catalytic activity. Characterisation of the proteomic response of C. mycophilum following exposure to B. velezensis culture filtrate revealed an increase in the abundance of a variety of proteins associated with stress response (ISWI chromatin-remodelling complex ATPase ISW2 (+24 fold), carboxypeptidase Y precursor (+3 fold) and calmodulin (+2 fold). There was also a decrease in the abundance of proteins associated with transcription (40 S ribosomal protein S30 (−26 fold), 40 S ribosomal protein S21 (−3 fold) and carbohydrate metabolism (l -xylulose reductase (−10 fold). The results presented here indicate that B. velezensis culture filtrate is capable of inhibiting the growth of C. mycophilum and inducing a stress response, thus indicating its potential to control this important pathogen of mushrooms. • Bacillus velezensis culture filtrate inhibits growth of Cladobotryum mycophilum. • Bacillus velezensis culture filtrate inhibits Cladobotryum mycophilum biomass accumulation. • C. mycophilum exposed to Bacillus velezensis culture filtrate shows oxidative stress response. • Bacillus velezensis culture filtrate contains range of lytic enzymes. • Bacillus velezensis culture filtrate may have potential to control cobweb. [ABSTRACT FROM AUTHOR]

Abstract The large pine weevil Hylobius abietis L. is a major forestry pest in 15 European countries, where it is a threat to 3.4 million hectares of forest. A cellular and proteomic analysis of the effect of culture filtrate of three entomopathogenic fungi (EPF) species on the immune system of H. abietis was performed. Injection with Metarhizium brunneum or Beauvaria bassiana culture filtrate facilitated a significantly increased yeast cell proliferation in larvae. Larvae co-injected with either Beauvaria caledonica or B. bassiana culture filtrate and Candida albicans showed significantly increased mortality. Together these results suggest that EPF culture filtrate has the potential to modulate the insect immune system allowing a subsequent pathogen to proliferate. Injection with EPF culture filtrate was shown to alter the abundance of protease inhibitors, detoxifing enzymes, antimicrobial peptides and proteins involved in reception/detection and development in H. abietis larvae. Larvae injected with B. caledonica culture filtrate displayed significant alterations in abundance of proteins involved in cellulolytic and other metabolic processes in their haemolymph proteome. Screening EPF for their ability to modulate the insect immune response represents a means of assessing EPF for use as biocontrol agents, particularly if the goal is to use them in combination with other control agents. Graphical abstract Image 1 Highlights • Injection with EPF filtrate facilitated an increased yeast cell density in larvae. • Co-injection with EPF and Candida albicans caused significantly increased mortality. • EPF can modulate insect immune system allowing proliferation of subsequent pathogens. • Injection with EPF culture filtrate significantly altered the haemolymph proteome. • Beauveria caledonica altered abundance of proteins involved in cellulolytic/metabolic processes. [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this study was to assess the accuracy of a virtual consultation (VC) process in determining treatment strategy for patients with malignant epidural spinal cord compression (MESCC). Methods and Materials: A prospective clinical database was maintained for patients with MESCC. A virtual consultation process (involving exchange of key predetermined clinical information and diagnostic imaging) facilitated rapid decision-making between oncologists and spinal surgeons. Diagnostic imaging was reviewed retrospectively (by R.R.) for surgical opinions in all patients. The primary outcome was the accuracy of virtual consultation opinion in predicting the final treatment recommendation. Results: After excluding 20 patients who were referred directly to the spinal surgeon, 125 patients were eligible for virtual consultation. Of the 46 patients who had a VC, surgery was recommended in 28 patients and actually given to 23. A retrospective review revealed that 5/79 patients who did not have a VC would have been considered surgical candidates. The overall accuracy of the virtual consultation process was estimated at 92%. Conclusion: The VC process for MESCC patients provides a reliable means of arriving at a multidisciplinary opinion while minimizing patient transfer. This can potentially shorten treatment decision time and enhance clinical outcomes. [Copyright &y& Elsevier]

An abstract of the article "Cortical Cartography Revisited: A Frequency Perspective on the Functional Architecture of Visual Cortex," by Amit Basole and colleagues is presented.

Ergothioneine (EGT) is a histidine betaine derivative that exhibits antioxidant action in humans. EGT is primarily synthesized by fungal species and a number of bacterial species. A five-gene cluster (egtA, egtB, egtC, egtD &egtE) responsible for EGT production in Mycobacteria smegmatis has recently been identified. The first fungal biosynthetic EGT gene (NcEgt-1) has also been identified in Neurospora crassa. NcEgt-1 contains domains similar to those found in M. smegmatis egtB and egtD. EGT is biomembrane impermeable. Here we inferred the evolutionary history of the EGT cluster in prokaryotes as well as examining the phyletic distribution of Egt-1 in the fungal kingdom. A genomic survey of 2509 prokaryotes showed that the five-gene EGT cluster is only found in the Actinobacteria. Our survey identified more than 400 diverse prokaryotes that contain genetically linked orthologs of egtB and egtD. Phylogenetic analyses of Egt proteins show a complex evolutionary history and multiple incidences of horizontal gene transfer. Our analysis also identified two independent incidences of a fusion event of egtB and egtD in bacterial species. A genomic survey of over 100 fungal genomes shows that Egt-1 is found in all fungal phyla, except species that belong to the Saccharomycotina subphylum. This analysis provides a comprehensive analysis of the distribution of the key genes involved in the synthesis of EGT in prokaryotes and fungi. Our phylogenetic inferences illuminate the complex evolutionary history of the genes involved in EGT synthesis in prokaryotes. The potential to synthesize EGT is a fungal trait except for species belonging to the Saccharomycotina subphylum. [ABSTRACT FROM AUTHOR]

Abstract: The purpose of this study was to examine the effect of joint angle on the relationship between force and electromyogram (EMG) amplitude and median frequency, in the biceps, brachioradialis and triceps muscles. Surface EMG were measured at eight elbow angles, during isometric flexion and extension at force levels from 10% to 100% of maximum voluntary contraction (MVC). Joint angle had a significant effect on MVC force, but not on MVC EMG amplitude in all of the muscles examined. The median frequency of the biceps and triceps EMG decreased with increasing muscle length, possibly due to relative changes in electrode position or a decrease in muscle fibre diameter. The relationship between EMG amplitude and force, normalised with respect to its maximum force at each angle, did not vary with joint angle in the biceps or brachioradialis muscles over all angles, or in the triceps between 45° and 120° of flexion. These results suggest that the neural excitation level to each muscle is determined by the required percentage of available force rather than the absolute force required. It is, therefore, recommended that when using surface EMG to estimate muscle excitation, force should be normalised with respect to its maximum value at each angle. [Copyright &y& Elsevier]

Cultivation of Agaricus bisporus is a large horticultural industry for many countries worldwide, where a single variety is almost grown exclusively. Mushroom virus X (MVX), a complex of multiple positive-sense single stranded RNA (ss(+)RNA) viruses, is a major pathogen of typical A. bisporus crops. MVX can manifest a variety of symptoms in crops and is highly infective and difficult to eradicate once established in host mycelium. Currently our knowledge regarding the molecular response of A. bisporus fruit bodies to MVX infection is limited. In order to study the response of different A. bisporus strains with different susceptibilities to MVX, we designed a model system to evaluate the in-vitro transmission of viruses in A. bisporus hyphae over a time-course, at two crucial phases in the crop cycle. The symptom expression of MVX in these varieties and the transcriptomic and proteomic response of fruit bodies to MVX-infection were examined. Transmission studies revealed the high potential of MVX to spread to uninfected mycelium yet not into the fruit bodies of certain strains in a crop. MVX affected colour and quality of multiple fruit bodies. Gene expression is significantly altered in all strains and between times of inoculation in the crop. Genes related to stress responses displayed differential expression. Proteomic responses revealed restriction of cellular signalling and vesicle transport in infected fruit bodies. This in-depth analysis examining many factors relevant to MVX infection in different A. bisporus strains, will provide key insights into host responses for this commercially important food crop. • MVX affected colour and quality of multiple fruit bodies. • MVX significantly alters gene expression in all strains. • Transcription factors and stress response genes expression were altered. • Proteomic revealed restriction of cellular signalling and vesicle transport. [ABSTRACT FROM AUTHOR]

Hyphae of filamentous fungi undergo polar extension, bifurcation and hyphal fusion to form reticulating networks of mycelia. Hyphal fusion or anastomosis, a ubiquitous process among filamentous fungi, is a vital strategy for how fungi expand over their substrate and interact with or recognise self- and non-self hyphae of neighbouring mycelia in their environment. Morphological and genetic characterisation of anastomosis has been studied in many model fungal species, but little is known of the direct proteomic response of two interacting fungal isolates. Agaricus bisporus , the most widely cultivated edible mushroom crop worldwide, was used as an in vitro model to profile the proteomes of interacting cultures. The globally cultivated strain (A15) was paired with two distinct strains; a commercial hybrid strain and a wild isolate strain. Each co-culture presented a different interaction ranging from complete vegetative compatibility (self), lack of interactions, and antagonistic interactions. These incompatible strains are the focus of research into disease-resistance in commercial crops as the spread of intracellular pathogens, namely mycoviruses, is limited by the lack of interhyphal anastomosis. Unique proteomic responses were detected between all co-cultures. An array of cell wall modifying enzymes, plus fungal growth and morphogenesis proteins were found in significantly (P < 0.05) altered abundances. Nitrogen metabolism dominated in the intracellular proteome, with evidence of nitrogen starvation between competing, non-compatible cultures. Changes in key enzymes of A. bisporus morphogenesis were observed, particularly via increased abundance of glucanosyltransferase in competing interactions and certain chitinases in vegetative compatible interactions only. Carbohydrate-active enzyme arsenals are expanded in antagonistic interactions in A. bisporus. Pathways involved in carbohydrate metabolism and genetic information processing were higher in interacting cultures, most notably during self-recognition. New insights into the differential response of interacting strains of A. bisporus will enhance our understanding of potential barriers to viral transmission through vegetative incompatibility. Our results suggest that a differential proteomic response occurs between A. bisporus at strain-level and findings from this work may guide future proteomic investigation of fungal anastomosis. • Unique proteomic responses detected between all Agaricus bisporus strain co-cultures. • Cell wall, fungal growth and morphogenesis proteins are differentially abundant. • Nitrogen starvation evident between competing and non-compatible strains. • Carbohydrate-active enzymes are expanded in antagonistic A. bisporus interactions. [ABSTRACT FROM AUTHOR]

Background: Sudden infant death syndrome (SIDS) is a leading cause of postneonatal mortality. Genetic heart diseases (GHDs) underlie some cases of SIDS.Objectives: This study aimed to determine the spectrum and prevalence of GHD-associated mutations as a potential monogenic basis for SIDS.Methods: A cohort of 419 unrelated SIDS cases (257 male; average age 2.7 ± 1.9 months) underwent whole exome sequencing and a targeted analysis of 90 GHD-susceptibility genes. The yield of "potentially informative," ultra-rare variants (minor allele frequency <0.00005) in GHD-associated genes was assessed.Results: Overall, 53 of 419 (12.6%) SIDS cases had ≥1 "potentially informative," GHD-associated variant. The yield was 14.9% (21 of 141) for mixed-European ancestry cases and 11.5% (32 of 278) for European ancestry SIDS cases. Infants older than 4 months were more likely to host a "potentially informative" GHD-associated variant. There was significant overrepresentation of ultra-rare nonsynonymous variants in European SIDS cases (18 of 278 [6.5%]) versus European control subjects (30 of 973 [3.1%]; p = 0.013) when combining all 4 major cardiac channelopathy genes (KCNQ1, KCNH2, SCN5A, and RYR2). According to the American College of Medical Genetics guidelines, only 18 of 419 (4.3%) SIDS cases hosted a "pathogenic" or "likely pathogenic" variant.Conclusions: Less than 15% of more than 400 SIDS cases had a "potentially informative" variant in a GHD-susceptibility gene, predominantly in the 4- to 12-month age group. Only 4.3% of cases possessed immediately clinically actionable variants. Consistent with previous studies, ultra-rare, nonsynonymous variants within the major cardiac channelopathy-associated genes were overrepresented in SIDS cases in infants of European ethnicity. These findings have major implications for the investigation of SIDS cases and families. [ABSTRACT FROM AUTHOR]

Galleria mellonella is a well-established model species regularly employed in the study of the insect immune response at cellular and humoral levels to investigate fungal pathogenesis and biocontrol agents. A cellular and proteomic analysis of the effect of culture filtrate of three entomopathogenic fungi (EPF) species on the immune system of G. mellonella was performed . Treatment with Beauveria caledonica and Metarhizium anisopliae 96 h culture filtrate facilitated a significantly increased yeast cell density in larvae (3-fold and 3.8-fold, respectively). Larvae co-injected with either M. anisopliae or B. caledonica culture filtrate and Candida albicans showed significantly increased mortality. The same was not seen for larvae injected with Beauveria bassiana filtrate. Together these results suggest that B. caledonica and M. anisopliae filtrate are modulating the insect immune system allowing a subsequent pathogen to proliferate. B. caledonica and M. anisopliae culture filtrates impact upon the larval prophenoloxidase (ProPO) cascade (e.g. ProPO activating factor 3 and proPO activating enzyme 3 were increased in abundance relative to controls), while B. bassiana treated larvae displayed higher abundances of alpha-esterase when compared to control larvae (2.4-fold greater) and larvae treated with M. anisopliae and B. caledonica . Treatment with EPF culture filtrate had a significant effect on antimicrobial peptide abundances particularly in M. anisopliae treated larvae where cecropin-D precursor, hemolin and gloverin were differentially abundant in comparison to controls. Differences in proteomic profiles for different treatments may reflect or even partially explain the differences in their immunomodulatory potential. Screening EPF for their ability to modulate the insect immune response represents a means of assessing EPF for use as biocontrol agents, particularly if the goal is to use them in combination with other control agents. Additionally EPF represent a valuable resource pool in our search for natural products with insect immunomodulatory and biocontrol properties. [ABSTRACT FROM AUTHOR]

Loss of function mutations in CTNNB1 have been reported in individuals with intellectual disability [MIM #615075] associated with peripheral spasticity, microcephaly and central hypotonia, suggesting a recognisable phenotype associated with haploinsufficiency for this gene. Trio based whole exome sequencing via the Deciphering Developmental Disorders (DDD) study has identified eleven further individuals with de novo loss of function mutations in CTNNB1 . Here we report detailed phenotypic information on ten of these. We confirm the features that have been previously described and further delineate the skin and hair findings, including fair skin and fair and sparse hair with unusual patterning. [ABSTRACT FROM AUTHOR]

Neurodevelopmental disorder with visual defects and brain anomalies (NEDVIBA) is a recently described genetic condition caused by de novo missense HK1 variants. Phenotypic data is currently limited; only seven patients have been published to date. This descriptive case series of a further four patients with de novo missense HK1 variants, alongside integration of phenotypic data with the reported cases, aims to improve our understanding of the associated phenotype. We provide further evidence that de novo HK1 variants located within the regulatory-terminal domain and alpha helix are associated with neurological problems and visual problems. We highlight for the first time an association with a raised cerebrospinal fluid lactate and specific abnormalities to the basal ganglia on brain magnetic resonance imaging, as well as associated respiratory issues and swallowing/feeding difficulties. We propose that this distinctive neurodevelopmental phenotype could arise through disruption of the regulatory glucose-6-phosphate binding site and subsequent gain of function of HK1 within the brain. [ABSTRACT FROM AUTHOR]

We describe a novel recognizable phenotype characterized by anophthalmia, a distinctive skeletal dysplasia and intellectual disability. Radiographic anomalies include severe rhizomelic shortness of the limbs and abnormal joint formation. Recent exome studies showed that these characteristics are part of the phenotypic spectrum of MAB21L2 gene mutations which cause a range of structural eye malformations such as microphthalmia/anophthalmia and ocular coloboma. The two unrelated individuals described here in detail are heterozygous carriers of the same de novo missense mutation c.151C > T (p.Arg51Cys) in MAB21L2. [ABSTRACT FROM AUTHOR]

Pierre Robin sequence (PRS) is an aetiologically distinct subgroup of cleft palate. We aimed to define the critical genomic interval from five different 5q22-5q31 deletions associated with PRS or PRS-associated features and assess each gene within the region as a candidate for the PRS component of the phenotype. Clinical array-based comparative genome hybridisation (aCGH) data were used to define a 2.08 Mb minimum region of overlap among four de novo deletions and one mother-son inherited deletion associated with at least one component of PRS. Commonly associated anomalies were talipes equinovarus (TEV), finger contractures and crumpled ear helices. Expression analysis of the orthologous genes within the PRS critical region in embryonic mice showed that the strongest candidate genes were FBN2 and PHAX . Targeted aCGH of the critical region and sequencing of these genes in a cohort of 25 PRS patients revealed no plausible disease-causing mutations. In conclusion, deletion of ∼2 Mb on 5q23 region causes a clinically recognisable subtype of PRS. Haploinsufficiency for FBN2 accounts for the digital and auricular features. A possible critical region for TEV is distinct and telomeric to the PRS region. The molecular basis of PRS in these cases remains undetermined but haploinsufficiency for PHAX is a plausible mechanism. [ABSTRACT FROM AUTHOR]

Abstract: A computational investigation of proximal humeral fracture fixation has been conducted. Four devices were selected for the study; a locking plate, intramedullary nail (IM Nail), K-wires and a Bilboquet device. A 3D model of a humerus was created using a process of thresholding based on the grayscale values of a CT scan of an intact humerus. An idealised three part fracture was created in addition to removing a standard volume from the humeral head as a representation of bone voids that occur as a result of the injury. All finite element simulations conducted represent 90° arm abduction. Simulations were conducted to investigate the effect of filling this bone void with calcium phosphate cement for each device. The effect of constructing devices from carbon fibre polyetheretherketone (CFPEEK) was investigated. Simulations of cement reinforced devices predict greater stability for each device. The average unreinforced fracture line opening (FLO) is reduced by 48.5% for metallic devices with a lesser effect on composite devices with FLO reduced by 23.6%. Relative sliding (shear displacement) is also reduced between fracture fragments by an average of 58.34%. CFPEEK device simulations predict reduced stresses at the device–bone interface. [Copyright &y& Elsevier]

Purpose: To develop a cone-beam computed tomography (CT)–enabled one-step simulation-to-treatment process for the treatment of bone metastases. Methods and Materials: A three-phase prospective study was conducted. Patients requiring palliative radiotherapy to the spine, mediastinum, or abdomen/pelvis suitable for treatment with simple beam geometry (≤2 beams) were accrued. Phase A established the accuracy of cone-beam CT images for the purpose of gross tumor target volume (GTV) definition. Phase B evaluated the feasibility of implementing the cone-beam CT–enabled planning process at the treatment unit. Phase C evaluated the online cone-beam CT–enabled process for the planning and treatment of patients requiring radiotherapy for bone metastases. Results: Eighty-four patients participated in this study. Phase A (n = 9) established the adequacy of cone-beam CT images for target definition. Phase B (n = 45) established the quality of treatment plans to be adequate for clinical implementation for bone metastases. When the process was applied clinically in bone metastases (Phase C), the degree of overlap between planning computed tomography (PCT) and cone-beam CT for GTV and between PCT and cone-beam CT for treatment field was 82% ± 11% and 97% ± 4%, respectively. The oncologist’s decision to accept the plan under a time-pressured environment remained of high quality, with the cone-beam CT–generated treatment plan delivering at least 90% of the prescribed dose to 100% ± 0% of the cone-beam CT planning target volume (PTV). With the assumption that the PCT PTV is the gold-standard target, the cone-beam CT–generated treatment plan delivered at least 90% and at least 95% of dose to 98% ± 2% and 97% ± 5% of the PCT PTV, respectively. The mean time for the online planning and treatment process was 32.7 ± 4.0 minutes. Patient satisfaction was high, with a trend for superior satisfaction with the cone-beam CT–enabled process. Conclusions: The cone-beam CT–enabled palliative treatment process is feasible and is ready for clinical implementation for the treatment of bone metastases using simple beam geometry, providing a streamlined one-step process toward palliative radiotherapy. [Copyright &y& Elsevier]

Abstract: Patellofemoral (PF)-related pathologies, including joint laxity, patellar maltracking, cartilage degradation and anterior knee pain, affect nearly 25% of the population. Researchers have investigated the influence of articular geometry on kinematics and contact mechanics in order to gain insight into the etiology of these conditions. The purpose of the current study was to create a three-dimensional statistical shape model of the PF joint and to characterize relationships between PF shape and function (kinematics and contact mechanics). A statistical shape model of the patellar and femoral articular surfaces and their relative alignment was developed from magnetic resonance images. Using 15 shape parameters, the model characterized 97% of the variation in the training set. The first three shape modes primarily described variation in size, patella alta–baja and depth of the sulcus groove. A previously verified finite element model was used to predict kinematics and contact mechanics for each subject. Combining the shape and joint mechanics data, a statistical shape–function model was developed that established quantitative relations of how changes in the shape of the PF joint influence mechanics. The predictive capability of the shape–function model was evaluated by comparing statistical model and finite element predictions, resulting in kinematic root mean square errors of less than 3° and 2.5mm. The key results of the study are dually in the implementation of a novel approach linking statistical shape and finite element models and the relationships elucidated between PF articular geometry and mechanics. [Copyright &y& Elsevier]

Purpose: To examine the long-term outcomes of a randomized trial comparing short (4 months; Arm 1) and long (8 months; Arm 2) neoadjuvant hormonal therapy before radiotherapy for localized prostate cancer. Methods and Materials: Between 1997 and 2001, 276 patients were enrolled and the data from 261 were analyzed. The stratification risk factors were prostate-specific antigen level >20 ng/mL, Gleason score ≥7, and Stage T3 or more. The intermediate-risk stratum had one factor and the high-risk stratum had two or more. Staging was done from the bone scan and computed tomography findings. The primary endpoint was biochemical failure-free survival. Results: The median follow-up was 102 months. The overall survival, biochemical failure-free survival. and prostate cancer-specific survival did not differ significantly between the two treatment arms, overall or at 5 years. The cumulative probability of overall survival at 5 years was 90% (range, 87–92%) in Arm 1 and 83% (range, 80–86%) in Arm 2. The biochemical failure-free survival rate at 5 years was 66% (range, 62–71%) in Arm 1 and 63% (range, 58–67%) in Arm 2. Conclusion: No statistically significant difference was found in biochemical failure-free survival between 4 months and 8 months of neoadjuvant hormonal therapy before radiotherapy for localized prostate cancer. [ABSTRACT FROM AUTHOR]

Abstract: This paper examines the development of innovation in human tissue technologies as a form of regenerative medicine, firstly by applying ‘pollution ideas’ to contemporary trends in its risk regulation and to the processes of regulatory policy formation, and secondly by analysing the classificatory processes deployed in regulatory policy. The analysis draws upon data from fieldwork and documentary materials with a focus on the UK and EU (2002–05) and explores four arenas: governance and regulatory policy; commercialisation and the market; ‘evidentiality’ manifest in evidence-based policy; and publics’ and technology users’ values and ethics. The analysis suggests that there is a trend toward ‘purification’ across these arenas, both material and socio-political. A common process of partitioning is found in stakeholders’ attempts to define a clear terrain, which the field of tissue-engineered technology might occupy. We conclude that pollution ideas and partitioning processes are useful in understanding regulatory ordering and innovation in the emerging technological zone of human tissue engineering. [Copyright &y& Elsevier]

Agaricus bisporus is a commercial mushroom crop susceptible to a disease caused by a complex of viruses known collectively as mushroom virus X (MVX). Symptoms of MVX include bare patches and mushroom cap discolouration (browning) in the fruiting bodies, phenotypes associated with the viruses AbV6 and AbV16, respectively. Limited understanding exists of the localisation and mobilisation of these viruses within the mycelium of A. bisporus. To this end, a non-destructive fluorescence in situ hybridisation (FISH) method was developed for in situ targeting of AbV6 and AbV16 in A. bisporus mycelium. An MVX strain associated with the bare patch disease phenotype revealed predominantly high signal towards the growing edges of cultures when probed for AbV6, with a 'halo-effect' of high signal intensity around putative vacuoles. An MVX strain associated with the browning disease phenotype showed high signal intensities within reticulating networks of hyphae in a highly compartmentalised manner when probed for AbV16. Localisation of the two viruses in MVX-infected cultures appears independent, as both viruses were found in completely discrete areas of the mycelium in differential patterns. FISH detected low level presence of the two viruses, AbV6 and AbV16 in a number of cultures which had tested negative for the viruses by RT-PCR. This suggests that FISH may be more sensitive at detecting viruses at low levels than molecular methods. This study demonstrates that FISH is a powerful tool in the field of mycovirology. • The technique FISH was used to target viruses within the mycelium of Agaricus bisporus. • Hyphae was cultured, permeabilised and hybridised with FISH probes on a single platform for fluorescence microscopy. • Two distinct viruses from the MVX complex were localised within different areas of the mycelium of A. bisporus strains. • Low levels of virus were detected within non-MVX strain hyphae and high levels were observed in MVX strain hyphae. [ABSTRACT FROM AUTHOR]

The European house dust mite, Dermatophagoides pteronyssinus is a major source of airborne allergens worldwide and is found in half of European homes. Interactions between microbes and house dust mites (HDM) are considered important factors that allow them to persist in the home. Laboratory studies indicate the European HDM, D. pteronyssinus is a mycophagous mite, capable of utilising a variety of fungi for nutrients, however specific mycolytic digestive enzymes are unknown. Our previous work identified a number of putative glycosyl hydrolases present in the predicted proteome of D. pteronyssinus airmid and validated the expression of 42 of these. Of note, three GH16 proteins with predicted β-1,3 glucanase activity were found to be consistently present in the mite body and excretome. Here, we performed an extensive bioinformatic, proteomic and biochemical study to characterize three-novel β-1,3 glucanases from this medically important house dust mite. The genes encoding novel β-1,3 glucanases designated Glu1 , Glu2 and Glu3 were identified in D. pteronyssinus airmid, each exhibited more than 59% amino acid identity to one another. These enzymes are encoded by Glu genes present in a tri-gene cluster and protein homologs are found in other acari. The patchy phyletic distribution of Glu proteins means their evolutionary history remains elusive, however horizontal gene transfer cannot be completely excluded. Recombinant Glu1 and Glu2 exhibit hydrolytic activity toward laminarin, pachyman and barley glucan. Excreted β-1,3 glucanase activity was increased in response to D. pteronyssinus airmid feeding on baker's yeast. Active β-1,3 glucanases are expressed and excreted in the faeces of D. pteronyssinus airmid indicating they are digestive enzymes capable of breaking down β-1,3 glucans of fungi present in house dust. Image 1 • β-1,3 glucanases found in HDMs and other Acari species, specifically those within the Acariformes order. • Mycophagous HDM, D. pteronyssinus , increases excretion of β-1,3 glucanases in response to Saccharomyces cerevisiae diet. • β-1,3 glucanases excreted by D. pteronyssinus are found in house dust. [ABSTRACT FROM AUTHOR]