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This paper presents a dynamic finite element method (FEM) model of a commercial displacement-controlled total knee replacement (TKR) wear tester. The first goal of the study was to validate the model, which included both the wear tester and the TKR components. Convergence simulations and experimental testing were performed. These included a novel experimental determination of the coefficient of friction and an evaluation of predicted joint contact areas by comparing simulation results with experimental data collected using pressure-sensitive film. The second goal of this study was to develop a procedure for implementing force-based testing protocols on a displacement-controlled TKR wear tester. A standard force-based cyclic wear-testing protocol was simulated using the FEM model and resulting displacement waveforms were extracted. These were used as control inputs to the physical wear tester and an experimental wear test was performed. Reaction loads on the tibial components were measured and compared with the simulated results. The model was capable of accurately predicting the tibial loads throughout the test cycle, verifying the model's contact mechanics. The study demonstrated the use of computational modelling to convert a force-based testing protocol into displacement-based control parameters for use in a displacement-controlled mechanical testing system. [ABSTRACT FROM AUTHOR]

Background: Patellofemoral pain syndrome is a prevalent condition in young people. While it is widely believed that abnormal patellar tracking plays a role in the development of patellofemoral pain syndrome, this link has not been established. The purpose of this cross-sectional case-control study was to test the hypothesis that patterns of patellar spin, tilt, and lateral translation make it possible to distinguish individuals with patellofemoral pain syndrome and clinical evidence of patellar malalignment from those with patellofemoral pain syndrome and no clinical evidence of malalignment and from individuals with no knee problems.Methods: Three-dimensional patellofemoral joint kinematics in one knee of each of sixty volunteers (twenty in each group described above) were assessed with use of a new, validated magnetic resonance imaging-based method. Static low-resolution scans of the loaded knee were acquired at five different angles of knee flexion (ranging between -4 degrees and 60 degrees). High-resolution geometric models of the patella, femur, and tibia and associated coordinate axes were registered to the bone positions on the low-resolution scans to determine the patellar motion as a function of knee flexion angle. Hierarchical modeling was used to identify group differences in patterns of patellar spin, tilt, and lateral translation.Results: No differences in the overall pattern of patellar motion were observed among groups (p>0.08 for all global maximum likelihood ratio tests). Features of patellar spin and tilt patterns varied greatly between subjects across all three groups, and no significant group differences were detected. At 19 degrees of knee flexion, the patellae in the group with patellofemoral pain and clinical evidence of malalignment were positioned an average of 2.25 mm more laterally than the patellae in the control group, and this difference was marginally significant (p=0.049). Other features of the pattern of lateral translation did not differ, and large overlaps in values were observed across all groups.Conclusions: It cannot be determined from our cross-sectional study whether the more lateral position of the patella in the group with clinical evidence of malalignment preceded or followed the onset of symptoms. It is clear from the data that an individual with patellofemoral pain syndrome cannot be distinguished from a control subject by examining patterns of spin, tilt, or lateral translation of the patella, even when clinical evidence of mechanical abnormality was observed. [ABSTRACT FROM AUTHOR]

En este estudio, se buscó investigar las concepciones de educadoras de guarderías públicas y privadas sobre desarrollo infantil. De ese modo, se aplicó una entrevista semiestructurada a 24 educadoras. Los datos fueron analizados a partir de la técnica de análisis de contenido categorial temático. De modo general, se verificó que, en relación a las concepciones sobre el desarrollo infantil, predominó, en los dos grupos de educadoras, la subclasificación Desarrollo por etapas. Esas educadoras destacaron el Ambiente socio-familiar desestructurado y el Despreparo del educador como siendo los principales aspectos desfavorables a la promoción del desarrollo infantil y caracterizan el desarrollo de los niños, cuando inseridos en la rutina de la guardería, como siendo positivo. Nuestro objetivo es, con este estudio, ofrecer subsidios para el planeamiento de intervenciones con vistas a promover el desarrollo infantil.

A computer-assisted technique using computed tomography and three-dimensional imaging for the localization and excision of osteoid osteoma allows minimal bone resection, shortened hospital stay, and prompt weight bearing. [ABSTRACT FROM AUTHOR]

Os profissionais da enfermagem têm se responsabilizado pela "arte" de cuidar, sendo eles os que dedicam a maior parte do seu tempo ao usuário. Neste estudo, objetivou-se compreender a relação entre auxiliar de enfermagem e usuário através das representações sociais do ato de tocar, construídas por esses profissionais no contexto do Programa Saúde da Família (PSF). Trata-se de uma pesquisa quali-quantitativa com 25 profissionais. Os instrumentos utilizados foram: associação livre de palavras, entrevista e questionário. Evidenciou-se que o toque representa possibilidades de aproximação entre profissional-usuário, expressando-se na prática cotidiana, pois auxilia na cura do usuário e humaniza a relação.Los profesionales de la enfermería tienen se responsabilizado por la "arte" de cuidar, siendo los que dedican la mayor parte de su tiempo al usuario. El objetivo de este trabajo es comprender la relación entre auxiliar de enfermería-usuario por las representaciones sociales del acto de tocar construidas por esos profesionales en el Programa Salud de la Familia. Se trata de una investigación cuali-cuantitativa con 25 profesionales. Los instrumentos utilizados fueron: asociación libre de palabras, entrevista y cuestionario. Se evidenció que el toque representa posibilidades de aproximación entre profesional-usuario, expresándose en la práctica cotidiana, pues posibilita la cura del usuario y humaniza la relación.Nursing professionals have themselves held liable for the "art" of take care of the users and are the ones who dedicate the major part of their time to them. The aim of this study is to understand the relationship between the nursing assistant and the user expressed by the social representations of the touch act by these professionals in the Family Health Program (PSF) context. The qualitative-quantitative research involved 25 nursing assistants. The instruments utilized were: free word association, interview, and questionnaire. It was observed that the touch represents possibilities of approximation between the professional and the user, as it helps in the user’s cure and humanizes the relationship.

RESUMEN: Este artículo tiene por objetivo discutir acerca del proceso de concretización de las políticas de educación inclusiva en realidades mexicanas, tomando como objeto de análisis el trabajo del director escolar en tres instituciones de enseñanza común, ubicadas en el municipio de Xalapa/VER que cuentan con la inscripción de alumnos con discapacidad. La recolección de datos fue realizada mientras una misión de estudios internacionales en el municipio de Xalapa/VER y el análisis de los datos consideró la literatura del campo de la Gestión escolar y Educación Especial.

The respiratory syncytial virus (RSV) fusion protein (F) facilitates virus-cell membrane fusion, which is critical for viral entry, and cell-cell fusion. In contrast to many type I fusion proteins, RSV F must be proteolytically cleaved at two distinct sites to be fusogenic. Cleavage at both sites results in the release of a 27 amino-acid fragment, termed Pep27. We examined proteolytic processing and the role of Pep27 for RSV F from both RSV A2 and RSV B9320 laboratory-adapted strains, allowing important comparisons between A and B clade F proteins. F from both clades was cleaved at both sites, and pulse-chase analysis indicated that cleavage at both sites occurs early after synthesis, most likely within the secretory pathway. Mutation of either site to alter the furin recognition motif blocked cell-cell fusion activity. To assess the role of Pep27 in F processing and expression, we deleted the Pep27 fragment, but preserved the cleavage sites. Deletion of Pep27 reduced F surface expression and cell-cell fusion. Two conserved N-linked glycosylation sites within Pep 27 are present in both the RSV A2 and RSV B9320 F. Randomization of the Pep27 sequence, while conserving the two N-liked glycosylation sites, did not significantly change surface expression, and only modestly reduced cell-cell fusion. However, the disruption of either Pep27 glycosylation site reduced cell-cell fusion. This work clarifies the timing of RSV F proteolytic cleavage and offers insight into the crucial role the N-linked glycosylation sites within Pep27 play in the biological function of F.

Although evolution is driven by changes in how regulatory pathways control development, we know little about the molecular details underlying these transitions. The TRA-2 domain that mediates contact with TRA-1 is conserved in Caenorhabditis. By comparing the interaction of these proteins in two species, we identified a striking change in how sexual development is controlled. Identical mutations in this domain promote oogenesis in Caenorhabditis elegans but promote spermatogenesis in Caenorhabditis briggsae. Furthermore, the effects of these mutations involve the male-promoting gene fem-3 in C. elegans but are independent of fem-3 in C. briggsae. Finally, reciprocal mutations in these genes show that C. briggsae TRA-2 binds TRA-1 to prevent expression of spermatogenesis regulators. By contrast, in C. elegans TRA-1 sequesters TRA-2 in the germ line, allowing FEM-3 to initiate spermatogenesis. Thus, we propose that the flow of information within the sex determination pathway has switched directions during evolution. This result has important implications for how evolutionary change can occur., (Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution 2024.)

DNA evidence in sexual assault cases have proven increasingly difficult to obtain and analyse due to increased condom use. With more interest in alternatives to DNA evidence, prophylactic lubricants, spermicides and residues may be interesting prospects. Current interest in the analysis of prophylactic residues focuses on the evaluation and identification of lubricants and constituents, primarily through gas chromatography or Fourier transform infrared spectroscopy. Though cost-effective methods, extensive sample preparation and destructive modes of analysis remain an area for improvement. As a result, the focus has since shifted to ambient ionization methods that offer adequate sensitivity and reduced sample preparation. The Liquid Microjunction Surface Sampling Probe (LMJSSP) is a versatile ambient ionization source that employs a probe that supports a continuously flushing droplet that extracts analytes when placed in contact with a surface. The analytes are aspirated into the mass spectrometer with a Venturi pressure. In this work we use the LMJSSP to analyse the trace transfer of condom lubricant to different types of fabric (cotton, cotton-spandex, and denim). Furthermore, we examine the sensitivity and storage conditions for the direct analysis method on different swab types (cotton, silicone, and foam). Additionally, Principal Component Analysis (PCA) and Maximally Collapsing Metric Learning (MCML) are utilized for visualization of differentiability of commercially available condom brands including Durex™ and Trojan™, and product subtypes. The results present an interesting multi-disciplinary approach of using a direct liquid extraction ambient ionization technique and machine learning to improve the overall workflow for the analysis of lubricants, swabs and fabrics. Machine learning algorithms were able to differentiate between inherent differences of Durex™ and Trojan™ condoms., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Richard David Oleschuk reports equipment, drugs, or supplies was provided by AB SCIEX LLC., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Importance: Human metapneumovirus is an important respiratory pathogen that causes significant morbidity and mortality, particularly in the very young, the elderly, and the immunosuppressed. However, the molecular details of how this virus spreads to new target cells are unclear. This work provides important new information on the formation of filamentous structures that are consistent with virus particles and adds critical new insight into the structure of extensions between cells that form during infection. In addition, it demonstrates for the first time the movement of viral replication centers through these intercellular extensions, representing a new mode of direct cell-to-cell spread that may be applicable to other viral systems., Competing Interests: The authors declare no conflict of interest.

The Paramyxoviridae family includes established human pathogens such as measles virus, mumps virus, and the human parainfluenza viruses; highly lethal zoonotic pathogens such as Nipah virus; and a number of recently identified agents, such as Sosuga virus, which remain poorly understood. The high human-to-human transmission rate of paramyxoviruses such as measles virus, high case fatality rate associated with other family members such as Nipah virus, and the existence of poorly characterized zoonotic pathogens raise concern that known and unknown paramyxoviruses have significant pandemic potential. In this review, the general life cycle, taxonomic relationships, and viral pathogenesis are described for paramyxoviruses that cause both systemic and respiratory system-restricted infections. Next, key gaps in critical areas are presented, following detailed conversations with subject matter experts and based on the current literature. Finally, we present an assessment of potential prototype pathogen candidates that could be used as models to study this important virus family, including assessment of the strengths and weaknesses of each potential prototype., Competing Interests: Potential conflicts of interest. Both authors: No reported conflicts of interest. Both authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed., (© The Author(s) 2023. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Although nematode genetics was founded on the use of hermaphrodite genetics for studying animal development and behavior, there is a growing need to extend this work to male/female species. One of the most promising species is C. nigoni, because it is so closely related to the model hermaphroditic C. briggsae. We present methods for using CRISPR/Cas9 gene editing to create mutations, and techniques for balancing, maintaining and studying these mutations., Competing Interests: The authors declare that there are no conflicts of interest present., (Copyright: © 2023 by the authors.)

Human metapneumovirus (HMPV) is a negative-strand RNA virus that frequently causes respiratory tract infections in infants, the elderly, and the immunocompromised. A hallmark of HMPV infection is the formation of membraneless, liquid-like replication and transcription centers in the cytosol termed inclusion bodies (IBs). The HMPV phosphoprotein (P) and nucleoprotein (N) are the minimal viral proteins necessary to form IB-like structures, and both proteins are required for the viral polymerase to synthesize RNA during infection. HMPV P is a homotetramer with regions of intrinsic disorder and has several known and predicted phosphorylation sites of unknown function. In this study, we found that the P C-terminal intrinsically disordered domain (CTD) must be present to facilitate IB formation with HMPV N, while either the N-terminal intrinsically disordered domain or the central oligomerization domain was dispensable. Alanine substitution at a single tyrosine residue within the CTD abrogated IB formation and reduced coimmunoprecipitation with HMPV N. Mutations to C-terminal phosphorylation sites revealed a potential role for phosphorylation in regulating RNA synthesis and P binding partners within IBs. Phosphorylation mutations which reduced RNA synthesis in a reporter assay produced comparable results in a recombinant viral rescue system, measured as an inability to produce infectious viral particles with genomes containing these single P mutations. This work highlights the critical role HMPV P plays in facilitating a key step of the viral life cycle and reveals the potential role for phosphorylation in regulating the function of this significant viral protein. IMPORTANCE Human metapneumovirus (HMPV) infects global populations, with severe respiratory tract infections occurring in infants, the elderly, and the immunocompromised. There are currently no FDA-approved therapeutics available to prevent or treat HMPV infection. Therefore, understanding how HMPV replicates is vital for the identification of novel targets for therapeutic development. During HMPV infection, viral RNA synthesis proteins localize to membraneless structures called inclusion bodies (IBs), which are sites of genome replication and transcription. The HMPV phosphoprotein (P) is necessary for IBs to form and for the virus to synthesize RNA, but it is not known how this protein contributes to IB formation or if it is capable of regulating viral replication. We show that the C-terminal domain of P is the location of a molecular interaction driving IB formation and contains potential phosphorylation sites where amino acid charge regulates the function of the viral polymerase complex., Competing Interests: The authors declare no conflict of interest.

To learn if orthologous mutations are temperature-sensitive in related species, we studied four C. briggsae mutations orthologous to alleles of important C. elegans genes. Both Cel-glp-4(bn2) and Cbr-glp-4(v473) are temperature-sensitive, causing sterility at 25°C. By contrast, Cel-fog-1 ( q253) is strongly ts , but its ortholog Cbr-fog-1(v442) causes a loss-of-function at all temperatures. Finally, the C. elegans glp-1 alleles bn18 and e2141 are ts sterile. However, their C. briggsae orthologs, Cbr-glp-1(v429) and Cbr-glp-1(v438) respectively, are wild-type at all temperatures. Thus, a ts mutation in one species provides clues about how to design ts alleles in another, but all theoretical outcomes are possible., (Copyright: © 2022 by the authors.)

Severe acute respiratory syndrome coronavirus 2's (SARS-CoV-2) rapid global spread has posed a significant threat to human health, and similar outbreaks could occur in the future. Developing effective virus inactivation technologies is critical to preventing and overcoming pandemics. The infection of SARS-CoV-2 depends on the binding of the spike glycoprotein (S) receptor binding domain (RBD) to the host cellular surface receptor angiotensin-converting enzyme 2 (ACE2). If this interaction is disrupted, SARS-CoV-2 infection could be inhibited. Magnetic nanoparticle (MNP) dispersions exposed to an alternating magnetic field (AMF) possess the unique ability for magnetically mediated energy delivery (MagMED); this localized energy delivery and associated mechanical, chemical, and thermal effects are a possible technique for inactivating viruses. This study investigates the MNPs' effect on vesicular stomatitis virus pseudoparticles containing the SARS-CoV-2 S protein when exposed to AMF or a water bath (WB) with varying target steady-state temperatures (45, 50, and 55 °C) for different exposure times (5, 15, and 30 min). In comparison to WB exposures at the same temperatures, AMF exposures resulted in significantly greater inactivation in multiple cases. This is likely due to AMF-induced localized heating and rotation of MNPs. In brief, our findings demonstrate a potential strategy for combating the SARS-CoV-2 pandemic or future ones.

Viruses frequently carry out replication in specialized compartments within cells. The effect of these structures on virus replication is poorly understood. Recent research supports phase separation as a foundational principle for organization of cellular components with the potential to influence viral replication. In this review, phase separation is described in the context of formation of viral replication centers, with an emphasis on the nonsegmented negative-strand RNA viruses. Consideration is given to the interplay between phase separation and the critical processes of viral transcription and genome replication, and the role of these regions in pathogen-host interactions is discussed. Finally, critical questions that must be addressed to fully understand how phase separation influences viral replication and the viral life cycle are presented, along with information about new approaches that could be used to make important breakthroughs in this emerging field.

Human metapneumovirus (HMPV) inclusion bodies (IBs) are dynamic structures required for efficient viral replication and transcription. The minimum components needed to form IB-like structures in cells are the nucleoprotein (N) and the tetrameric phosphoprotein (P). HMPV P binds to the following two versions of the N protein in infected cells: N-terminal P residues interact with monomeric N (N 0 ) to maintain a pool of protein to encapsidate new RNA and C-terminal P residues interact with oligomeric, RNA-bound N (N-RNA). Recent work on other negative-strand viruses has suggested that IBs are, at least in part, liquid-like phase-separated membraneless organelles. Here, HMPV IBs in infected or transfected cells were shown to possess liquid organelle properties, such as fusion and fission. Recombinant versions of HMPV N and P proteins were purified to analyze the interactions required to drive phase separation in vitro . Purified HMPV P was shown to form liquid droplets in isolation. This observation is distinct from other viral systems that also form IBs. Partial removal of nucleic acid from purified P altered phase-separation dynamics, suggesting that nucleic acid interactions play a role in IB formation. HMPV P also recruits monomeric N (N 0 -P) and N-RNA to droplets in vitro . These findings suggest that HMPV P may also act as a scaffold protein to mediate multivalent interactions with monomeric and oligomeric N, as well as RNA, to promote phase separation of IBs. Together, these findings highlight an additional layer of regulation in HMPV replication by the viral P and N proteins. IMPORTANCE Human metapneumovirus (HMPV) is a leading cause of respiratory disease among children, immunocompromised individuals, and the elderly. Currently, no vaccines or antivirals are available for the treatment of HMPV infections. Cytoplasmic inclusion bodies (IBs), where HMPV replication and transcription occur, represent a promising target for the development of novel antivirals. The HMPV nucleoprotein (N) and phosphoprotein (P) are the minimal components needed for IB formation in eukaryotic cells. However, interactions that regulate the formation of these dynamic structures are poorly understood. Here, we showed that HMPV IBs possess the properties of liquid organelles and that purified HMPV P phase separates independently in vitro . Our work suggests that HMPV P phase-separation dynamics are altered by nucleic acid. We provide strong evidence that, unlike results reported from other viral systems, HMPV P alone can serve as a scaffold for multivalent interactions with monomeric (N 0 ) and oligomeric (N-RNA) HMPV N for IB formation.

Purpose: To determine if eccentric exercise was effective, safe and feasible in increasing function and quality of life in people with heart failure compared to usual care and a waitlist control group., Methods: A prospective, three-armed, parallel-design, assessor-blind, pilot randomised controlled trial with 1:1:1 allocation. Forty-seven participants (16 female; mean age 66 years) with mild to moderate heart failure were randomly allocated to either eccentric exercise, concentric exercise or a waitlist control group. Participants in the exercise groups completed twice-weekly exercise for eight weeks. Primary outcome was walking capacity. Secondary outcomes were quality of life, leg strength and fatigue. Outcomes were assessed at baseline, post intervention and three-month follow-up. Attendance, tolerability and adverse events were used to determine safety and feasibility., Results: Intention-to-treat analysis showed no differences between eccentric exercise and either concentric exercise or waitlist for any outcome. Per-protocol analysis found improvements identified by the Minnesota living with heart failure questionnaire were significantly greater post-intervention for eccentric exercise compared to concentric exercise (-17.99 units, 95% confidence interval -35.96 to -0.01). No major adverse events were reported., Conclusion: In this small trial, eccentric exercise did not demonstrate superior outcomes to concentric exercise or a waitlist control group., Clinical Trial Registration: The protocol for this trial was registered at clinicaltrials.gov, registration number: NCT02223624, registration date: 22 August 2014.IMPLICATIONS FOR REHABILITATIONRegular physical activity and referral to rehabilitation is recommended for people with chronic heart failure, however exercise can be challenging for this group.Eccentric exercise was safe and tolerable for participants with heart failure.Documentation of exercise progression is important to demonstrate a dose-response relationship.In this study there were no differences between groups who received eccentric exercise, concentric exercise or no exercise.

Background: Animal germ cells differentiate as sperm or as oocytes. These sexual fates are controlled by complex regulatory pathways to ensure that the proper gametes are made at the appropriate times., Summary: Nematodes like Caenorhabditis elegans and its close relatives are ideal models for studying how this regulation works, because the XX animals are self-fertile hermaphrodites that produce both sperm and oocytes. In these worms, germ cells use the same signal transduction pathway that functions in somatic cells. This pathway determines the activity of the transcription factor TRA-1, a Gli protein that can repress male genes. However, the pathway is extensively modified in germ cells, largely by the action of translational regulators like the PUF proteins. Many of these modifications play critical roles in allowing the XX hermaphrodites to make sperm in an otherwise female body. Finally, TRA-1 cooperates with chromatin regulators in the germ line to control the activity of fog-1 and fog-3, which are essential for spermatogenesis. FOG-1 and FOG-3 work together to determine germ cell fates by blocking the translation of oogenic transcripts., Key Messages: Although there is great diversity in how germ cell fates are controlled in other animals, many of the key nematode genes are conserved, and the critical role of translational regulators may be universal., (© 2022 S. Karger AG, Basel.)

Hendra virus (HeV) is a zoonotic enveloped member of the family Paramyoxviridae. To successfully infect a host cell, HeV utilizes two surface glycoproteins: the attachment (G) protein to bind, and the trimeric fusion (F) protein to merge the viral envelope with the membrane of the host cell. The transmembrane (TM) region of HeV F has been shown to have roles in F protein stability and the overall trimeric association of F. Previously, alanine scanning mutagenesis has been performed on the C-terminal end of the protein, revealing the importance of β-branched residues in this region. Additionally, residues S490 and Y498 have been demonstrated to be important for F protein endocytosis, needed for the proteolytic processing of F required for fusion. To complete the analysis of the HeV F TM, we performed alanine scanning mutagenesis to explore the residues in the N-terminus of this region (residues 487-506). In addition to confirming the critical roles for S490 and Y498, we demonstrate that mutations at residues M491 and L492 alter F protein function, suggesting a role for these residues in the fusion process.

Purpose: Intraoperative assessment of surgical margins is important for reducing the rate of revisions in breast conserving surgery for palpable malignant tumors. The hypothesis was that metabolomics methods, based on mass spectrometry, could find patterns of relative abundances of molecules that distinguish clusters of benign tissue and cancer in surgical resections., Methods: Excisions from 8 patients were used to acquire 112,317 mass spectrometry signals by desorption electrospray ionization. A process of nonnegative matrix factorization and graph decomposition produced clusters that were approximated as affine spaces. Each signal's distance to the affine space of a cluster was used to visualize the clustering., Results: The distance maps were superior to binary clustering in identifying cancer regions. They were particularly effective at finding cancer regions that were discontinuously distributed within benign tissue., Conclusions: Desorption electrospray ionization mass spectrometry, which has been shown to be useful intraoperatively, can acquire signals that distinguish malignant from benign breast tissue in surgically excised tumors. The method may be suitable for real-time surgical decisions based on cancer margins.

There is increasing evidence implicating altered NMDA-receptor function in autism spectrum disorder (ASD). To investigate potential alterations in NMDA-dependent cortical plasticity in ASD, we examined the effect of visual high-frequency stimulation (HFS) on changes in plasticity in the visual cortex, measured by persistent changes in visual evoked potentials (VEPs), in individuals with ASD (n = 16) and neurotypical controls (NT; n = 15). VEPs were elicited by a checkerboard circle (0.83 Hz, 2-min blocks) at baseline and at 2, 4, and 20 min following exposure to HFS (8.87 Hz, 2 min), previously shown to induce LTP-like changes in the visual cortex. Difference waves were created by subtracting VEPs measured at baseline from each Post-HFS measure, and group differences assessed. We found that HFS resulted in reduced short-term potentiation of VEPs in ASD compared to NT participants. Thus, whilst ASD participants showed significant potentiation of the VEP immediately after HFS, this enhancement was not maintained, and only persisted into the second post-HFS assessment block in NT participants. Notably, ASD individuals who self-reported being more sensitive to visual stimuli showed greater shorter-term potentiation following visual HFS. Critically, there were no group differences in degree of neural entrainment to the visual HFS, or in attentional vigilance and task performance. These findings suggests that visual cortical plasticity is atypical in ASD, results consistent with reported altered NMDA receptor function in ASD., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Changing how hormones regulate development can influence the complex trade-offs involved in the competition to survive and reproduce. A new report identifies a molecular variant of eak-3 that trades lower growth rates for the ability to survive sudden temperature extremes., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Respiratory syncytial virus (RSV) and human metapneumovirus (HMPV) are two of the leading causes of respiratory infections in children and elderly and immunocompromised patients worldwide. There is no approved treatment for HMPV and only one prophylactic treatment against RSV, palivizumab, for high-risk infants. Better understanding of the viral lifecycles in a more relevant model system may help identify novel therapeutic targets. By utilizing three-dimensional (3-D) human airway tissues to examine viral infection in a physiologically relevant model system, we showed that RSV infects and spreads more efficiently than HMPV, with the latter requiring higher multiplicities of infection (MOIs) to yield similar levels of infection. Apical ciliated cells were the target for both viruses, but RSV apical release was significantly more efficient than HMPV. In RSV- or HMPV-infected cells, cytosolic inclusion bodies containing the nucleoprotein, phosphoprotein, and respective viral genomic RNA were clearly observed in human airway epithelial (HAE) culture. In HMPV-infected cells, actin-based filamentous extensions were more common (35.8%) than those found in RSV-infected cells (4.4%). Interestingly, neither RSV nor HMPV formed syncytia in HAE tissues. Palivizumab and nirsevimab effectively inhibited entry and spread of RSV in HAE tissues, with nirsevimab displaying significantly higher potency than palivizumab. In contrast, 54G10 completely inhibited HMPV entry but only modestly reduced viral spread, suggesting HMPV may use alternative mechanisms for spread. These results represent the first comparative analysis of infection by the two pneumoviruses in a physiologically relevant model, demonstrating an interesting dichotomy in the mechanisms of infection, spread, and consequent inhibition of the viral lifecycles by neutralizing monoclonal antibodies. IMPORTANCE Respiratory syncytial virus and human metapneumovirus are leading causes of respiratory illness worldwide, but limited treatment options are available. To better target these viruses, we examined key aspects of the viral life cycle in three-dimensional (3-D) human airway tissues. Both viruses establish efficient infection through the apical surface, but efficient spread and apical release were seen for respiratory syncytial virus (RSV) but not human metapneumovirus (HMPV). Both viruses form inclusion bodies, minimally composed of nucleoprotein (N), phosphoprotein (P), and viral RNA (vRNA), indicating that these structures are critical for replication in this more physiological model. HMPV formed significantly more long, filamentous actin-based extensions in human airway epithelial (HAE) tissues than RSV, suggesting HMPV may promote cell-to-cell spread via these extensions. Lastly, RSV entry and spread were fully inhibited by neutralizing antibodies palivizumab and the novel nirsevimab. In contrast, while HMPV entry was fully inhibited by 54G10, a neutralizing antibody, spread was only modestly reduced, further supporting a cell-to-cell spread mechanism., (Copyright © 2020 American Society for Microbiology.)

In nematodes, TRA-1 represses the transcription of genes involved in male differentiation, allowing XX animals to undergo normal hermaphrodite development. New reports show that this transcription factor also acts in XO males, to control the differentiation of many neurons., (Copyright © 2020 Elsevier Inc. All rights reserved.)

During craniotomies, a portion of the calvarium or skull is removed to gain access to the intracranial space. When it is not possible to re-implant the flap, surgeons may repair the defect intraoperatively or at a later date. With larger defects being more difficult to repair intraoperatively, we investigated a method for the creation of patient-specific moulds for ad hoc bone flap reconstruction using rapid prototyping. Patient-specific moulds were created based on light scanned models of the defect, using custom software and rapid prototyping. Polymethylmethacrylate bone implants were created for three retrospective craniotomy cases and evaluated based on original flap and skull reconstruction accuracy. Bone implants created using our moulding method reconstruct the original flap and skull with an average reconstruction accuracy of 0.82 and 1.3 mm, respectively. Average skull reconstruction accuracy obtained by surgeons performing freehand implant reconstruction was 1.49 mm. Time needed to generate moulds was between 2 h and 45 min and 6 h and 20 min. Improvements to current printing technology will make this procedure technically feasible for future cranial procedures.

Initiation of host cell infection by an enveloped virus requires a viral-to-host cell membrane fusion event. This event is mediated by at least one viral transmembrane glycoprotein, termed the fusion protein, which is a key therapeutic target. Viral fusion proteins have been studied for decades, and numerous critical insights into their function have been elucidated. However, the transmembrane region remains one of the most poorly understood facets of these proteins. In the past ten years, the field has made significant advances in understanding the role of the membrane-spanning region of viral fusion proteins. We summarize developments made in the past decade that have contributed to the understanding of the transmembrane region of viral fusion proteins, highlighting not only their critical role in the membrane fusion process, but further demonstrating their involvement in several aspects of the viral lifecycle., Competing Interests: The authors report no conflict of interest.

Objectives: Osteophytes are common anatomical signs of advanced osteoarthritis. It remains unclear whether they develop from physio-molecular, and/or mechanical stimuli. This study examined the effects of mechanical impact on the knee joint periosteum leading to osteophyte formation., Design: Eighteen mature rats received one single impact load of 53 N (30 MPa) to the periosteum of the experimental medial femoral condyles. Contralateral knees were used as controls. Animals were sacrificed at 24 h, 3, 6 and 9 weeks post-impact. Distal femurs were harvested and prepared for histology. Hematoxylin and Eosin, and Masson's trichrome stained slides were examined by light microscopy. Nuclear density was quantified to assess the tissue reaction., Results: 24 h: The synovium membrane, fibrous and cambium periosteum were damaged. Blood infiltration pooled in the impacted medial collateral ligament (MCL) region. Week 3: A cartilaginous tissue spur, chondrophyte, was found in every rat at the impacted site of the MCL. Chondrophytes were composed of fibrocartilage and cartilage matrix, with signs of cartilage mineralization and remodelling activity. Week 6: Chondrophytes presented signs of more advanced mineralisation, recognized as osteophytes. Week 9: Osteophytes appeared to be more mineralized with almost no cartilage tissue., Conclusions: Osteophytes can be induced with a single mechanical impact applied to the periosteum in rat knees. These data indicate that a moderate trauma to the periosteal layer of the joint may play a role in osteophyte development., (Copyright © 2020 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

The paramyxoviruses Hendra virus (HeV) and parainfluenza virus 5 (PIV5) require the fusion (F) protein to efficiently infect cells. For fusion to occur, F undergoes dramatic, essentially irreversible conformational changes to merge the viral and cell membranes into a continuous bilayer. Recently, a transmembrane (TM) domain leucine/isoleucine (L/I) zipper was shown to be critical in maintaining the expression, stability and pre-fusion conformation of HeV F, allowing for fine-tuned timing of membrane fusion. To analyse the effect of the TM domain L/I zipper in another paramyxovirus, we created alanine mutations to the TM domain of PIV5 F, a paramyxovirus model system. Our data show that while the PIV5 F TM L/I zipper does not significantly affect total expression and only modestly affects surface expression and pre-fusion stability, it is critical for fusogenic activity. These results suggest that the roles of TM L/I zipper motifs differ among members of the family Paramyxoviridae .

Viruses possessing class I fusion proteins require proteolytic activation by host cell proteases to mediate fusion with the host cell membrane. The mammalian SPINT2 gene encodes a protease inhibitor that targets trypsin-like serine proteases. Here we show the protease inhibitor, SPINT2, restricts cleavage-activation efficiently for a range of influenza viruses and for human metapneumovirus (HMPV). SPINT2 treatment resulted in the cleavage and fusion inhibition of full-length influenza A/CA/04/09 (H1N1) HA, A/Aichi/68 (H3N2) HA, A/Shanghai/2/2013 (H7N9) HA and HMPV F when activated by trypsin, recombinant matriptase or KLK5. We also demonstrate that SPINT2 was able to reduce viral growth of influenza A/CA/04/09 H1N1 and A/X31 H3N2 in cell culture by inhibiting matriptase or TMPRSS2. Moreover, inhibition efficacy did not differ whether SPINT2 was added at the time of infection or 24 h post-infection. Our data suggest that the SPINT2 inhibitor has a strong potential to serve as a novel broad-spectrum antiviral., 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 © 2020 Elsevier Inc. All rights reserved.)

A critical step in the life cycle of a virus is spread to a new target cell, which generally involves the release of new viral particles from the infected cell which can then initiate infection in the next target cell. While cell-free viral particles released into the extracellular environment are necessary for long distance spread, there are disadvantages to this mechanism. These include the presence of immune system components, the low success rate of infection by single particles, and the relative fragility of viral particles in the environment. Several mechanisms of direct cell-to-cell spread have been reported for animal viruses which would avoid the issues associated with cell-free particles. A number of viruses can utilize several different mechanisms of direct cell-to-cell spread, but our understanding of the differential usage by these pathogens is modest. Although the mechanisms of cell-to-cell spread differ among viruses, there is a common exploitation of key pathways and components of the cellular cytoskeleton. Remarkably, some of the viral mechanisms of cell-to-cell spread are surprisingly similar to those used by bacteria. Here we summarize the current knowledge of the conventional and non-conventional mechanisms of viral spread, the common methods used to detect viral spread, and the impact that these mechanisms can have on viral pathogenesis., (© 2020 Elsevier Inc. All rights reserved.)

Hendra virus (HeV) is a zoonotic paramyxovirus that utilizes a trimeric fusion (F) protein within its lipid bilayer to mediate membrane merger with a cell membrane for entry. Previous HeV F studies showed that transmembrane domain (TMD) interactions are important for stabilizing the prefusion conformation of the protein prior to triggering. Thus, the current model for HeV F fusion suggests that modulation of TMD interactions is critical for initiation and completion of conformational changes that drive membrane fusion. HeV F constructs (T483C/V484C, V484C/N485C, and N485C/P486C) were generated with double cysteine substitutions near the N-terminal region of the TMD to study the effect of altered flexibility in this region. Oligomeric analysis showed that the double cysteine substitutions successfully promoted intersubunit disulfide bond formation in HeV F. Subsequent fusion assays indicated that the introduction of disulfide bonds in the mutants prohibited fusion events. Further testing confirmed that T483C/V484C and V484C/N485C were expressed at the cell surface at levels that would allow for fusion. Attempts to restore fusion with a reducing agent were unsuccessful, suggesting that the introduced disulfide bonds were likely buried in the membrane. Conformational analysis showed that T483C/V484C and V484C/N485C were able to bind a prefusion conformation-specific antibody prior to cell disruption, indicating that the introduced disulfide bonds did not significantly affect protein folding. This study is the first to report that TMD dissociation is required for HeV F fusogenic activity and strengthens our model for HeV fusion. IMPORTANCE The paramyxovirus Hendra virus (HeV) causes severe respiratory illness and encephalitis in humans. To develop therapeutics for HeV and related viral infections, further studies are needed to understand the mechanisms underlying paramyxovirus fusion events. Knowledge gained in studies of the HeV fusion (F) protein may be applicable to a broad span of enveloped viruses. In this study, we demonstrate that disulfide bonds introduced between the HeV F transmembrane domains (TMDs) block fusion. Depending on the location of these disulfide bonds, HeV F can still fold properly and bind a prefusion conformation-specific antibody prior to cell disruption. These findings support our current model for HeV membrane fusion and expand our knowledge of the TMD and its role in HeV F stability and fusion promotion., (Copyright © 2019 American Society for Microbiology.)

Genes can encode multiple isoforms, broadening their functions and providing a molecular substrate to evolve phenotypic diversity. Evolution of isoform function is a potential route to adapt to new environments. Here we show that de novo, beneficial alleles in the nurf-1 gene became fixed in two laboratory lineages of C. elegans after isolation from the wild in 1951, before methods of cryopreservation were developed. nurf-1 encodes an ortholog of BPTF, a large (>300 kD) multidomain subunit of the NURF chromatin remodeling complex. Using CRISPR-Cas9 genome editing and transgenic rescue, we demonstrate that in C. elegans , nurf-1 has split into two, largely non-overlapping isoforms (NURF-1.D and NURF-1.B, which we call Yin and Yang, respectively) that share only two of 26 exons. Both isoforms are essential for normal gametogenesis but have opposite effects on male/female gamete differentiation. Reproduction in hermaphrodites, which involves production of both sperm and oocytes, requires a balance of these opposing Yin and Yang isoforms. Transgenic rescue and genetic position of the fixed mutations suggest that different isoforms are modified in each laboratory strain. In a related clade of Caenorhabditis nematodes, the shared exons have duplicated, resulting in the split of the Yin and Yang isoforms into separate genes, each containing approximately 200 amino acids of duplicated sequence that has undergone accelerated protein evolution following the duplication. Associated with this duplication event is the loss of two additional nurf-1 transcripts, including the long-form transcript and a newly identified, highly expressed transcript encoded by the duplicated exons. We propose these lost transcripts are non-functional side products necessary to transcribe the Yin and Yang transcripts in the same cells. Our work demonstrates how gene sharing, through the production of multiple isoforms, can precede the creation of new, independent genes., Competing Interests: WX, LL, YZ, LS, IF, JM, RE, PM No competing interests declared, (© 2019, Xu et al.)

Enveloped viruses utilize surface glycoproteins to bind and fuse with a target cell membrane. The zoonotic Hendra virus (HeV), a member of the family Paramyxoviridae , utilizes the attachment protein (G) and the fusion protein (F) to perform these critical functions. Upon triggering, the trimeric F protein undergoes a large, irreversible conformation change to drive membrane fusion. Previously, we have shown that the transmembrane (TM) domain of the F protein, separate from the rest of the protein, is present in a monomer-trimer equilibrium. This TM-TM association contributes to the stability of the prefusion form of the protein, supporting a role for TM-TM interactions in the control of F protein conformational changes. To determine the impact of disrupting TM-TM interactions, constructs expressing the HeV F TM with limited flanking sequences were synthesized. Coexpression of these constructs with HeV F resulted in dramatic reductions in the stability of F protein expression and fusion activity. In contrast, no effects were observed when the HeV F TM constructs were coexpressed with the nonhomologous parainfluenza virus 5 (PIV5) fusion protein, indicating a requirement for specific interactions. To further examine this, a TM peptide homologous to the PIV5 F TM domain was synthesized. Addition of the peptide prior to infection inhibited infection with PIV5 but did not significantly affect infection with human metapneumovirus, a related virus. These results indicate that targeted disruption of TM-TM interactions significantly impact viral fusion protein stability and function, presenting these interactions as a novel target for antiviral development. IMPORTANCE Enveloped viruses require virus-cell membrane fusion to release the viral genome and replicate. The viral fusion protein triggers from the pre- to the postfusion conformation, an essentially irreversible change, to drive membrane fusion. We found that small proteins containing the TM and a limited flanking region homologous to the fusion protein of the zoonotic Hendra virus reduced protein expression and fusion activity. The introduction of exogenous TM peptides may displace a TM domain, disrupting native TM-TM interactions and globally destabilizing the fusion protein. Supporting this hypothesis, we showed that a sequence-specific transmembrane peptide dramatically reduced viral infection in another enveloped virus model, suggesting a broader inhibitory mechanism. Viral fusion protein TM-TM interactions are important for protein function, and disruption of these interactions dramatically reduces protein stability., (Copyright © 2019 American Society for Microbiology.)

Background: Long-term function and survival of reverse shoulder arthroplasties (RSAs) are reliant on component positioning and fixation. Conventional postoperative analysis is performed using plain radiographs or 2-dimensional (2D) computed tomography (CT) images. Although 3-dimensional (3D) CT would be preferred, its use is limited by metal artifacts. This study proposes a new 3D CT method for postoperative RSA evaluation and compares its interobserver reliability with conventional methods., Materials and Methods: Preoperative and postoperative CT scans, as well as postoperative radiographs, were obtained from 18 patients who underwent RSA implantation; the scapula, implant, and screws were reconstructed as 3D CT models. The postoperative 3D scapula and implant were imported into preoperative coordinates and matched to the preoperative scapula. Standardized scapula coordinates were defined, in which the glenoid baseplate version and inclination angle were measured. The percentage of screw volume in bone was measured from a Boolean intersection operation between the preoperative scapula and screw models. Four independent reviewers performed the measurements using 3D CT and conventional 2D methods. Intraclass correlation coefficients (ICCs) were used to compare the reliability of the methods., Results: The 3D CT method showed excellent reliability (ICC > 0.75) in baseplate inclination (ICC = 0.92), version (ICC = 0.97), and screw volume in bone (ICC = 0.99). Conventional 2D methods demonstrated poor reliability (ICC < 0.4). For radiographs, inclination showed poor reliability (ICC = 0.09) and the screw percentage in bone showed fair reliability (ICC = 0.54). Version was not measured with plain radiographs. For 2D CT slice measurements, inclination showed poor reliability (ICC = 0.02), version showed excellent reliability (ICC = 0.81), and the screw percentage in bone showed poor reliability (ICC = 0.28)., Conclusion: The new 3D CT-based method for evaluating RSA glenoid implant positioning and screw volume in bone showed excellent reliability and overcame the metal-artifact limitation of postoperative CT and 3D CT reconstruction., (© 2019 The Authors.)

Purpose: Minimally invasive beating-heart surgery is currently performed using endoscopes and without navigation. Registration of intraoperative ultrasound to a preoperative cardiac CT scan is a valuable step toward image-guided navigation., Methods: The registration was achieved by first extracting a representative point set from each ultrasound image in the sequence using a deformable registration. A template shape representing the cardiac chambers was deformed through a hierarchy of affine transformations to match each ultrasound image using a generalized expectation maximization algorithm. These extracted point sets were matched to the CT by exhaustively searching over a large number of precomputed slices of 3D geometry. The result is a similarity transformation mapping the intraoperative ultrasound to preoperative CT., Results: Complete data sets were acquired for four patients. Transesophageal echocardiography ultrasound sequences were deformably registered to a model of oriented points with a mean error of 2.3 mm. Ultrasound and CT scans were registered to a mean of 3 mm, which is comparable to the error of 2.8 mm expected by merging ultrasound registration with uncertainty of cardiac CT., Conclusion: The proposed algorithm registered 3D CT with dynamic 2D intraoperative imaging. The algorithm aligned the images in both space and time, needing neither dynamic CT imaging nor intraoperative electrocardiograms. The accuracy was sufficient for navigation in thoracoscopically guided beating-heart surgery.

The human metapneumovirus (HMPV) fusion protein (F) mediates fusion of the viral envelope and cellular membranes to establish infection. HMPV F from some, but not all, viral strains promotes fusion only after exposure to low pH. Previous studies have identified several key residues involved in low pH triggering, including H435 and a proposed requirement for glycine at position 294. We analyzed the different levels of fusion activity, protein expression and cleavage of three HMPV F proteins not previously examined. Interestingly, low pH-triggered fusion in the absence of G294 was identified in one F protein, while a novel histidine residue (H434) was identified that enhanced low pH promoted fusion in another. The third F protein failed to promote cell-to-cell fusion, suggesting other requirements for F protein triggering. Our results demonstrate HMPV F triggering is more complex than previously described and suggest a more intricate mechanism for fusion protein function and activation., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The adverse effects formaldehyde fixation has on tissues both gross anatomically and histologically are well documented. Consequently, researchers are seeking alternative embalming techniques that better preserve in vivo characteristics of tissues. Phenol-based embalming is one method that has shown promise in its ability to adequately preserve the in vivo qualities of tissues through preliminary explorations at the gross anatomical level. The literature on phenol-based embalming is currently scarce, especially with regard to its effects on tissues at the microscopic level. For the current study we aimed to document the histologic effects of a formaldehyde-free phenol-based embalming solution on neural tissue, with the hope of providing novel insight into the effects of soft-embalming on tissues at the microscopic level. Cerebral and cerebellar tissue obtained from porcine brains was fixed in phenol- and formaldehyde-based fixatives; the latter served as a control. Fixed samples were processed for histological analysis. The phenol-based embalming solution provided excellent preservation of the cerebral and cerebellar tissue morphology. Of note was the decrease in separation artifact seen in both tissue types relative to the control tissue, as well as anomalous circular artifacts in the white matter. The results of this study indicate that the phenol-based embalming solution preserves neural tissue at the histological level, perhaps superiorly in many aspects when compared to the formaldehyde-fixed samples. Further investigations of both gross anatomy and histology are recommended on the basis of these promising new findings to determine its potential utilities within research and education. Clin. Anat. 32:224-230, 2019. © 2018 Wiley Periodicals, Inc., (© 2018 Wiley Periodicals, Inc.)

Objectives: The aim of this study is to evaluate the hepatoprotective effect of Lasianthera africana (Icacinaceae) against isoniazid (INH) and rifampicin (RIF)-induced liver damage in rats., Methods: The hepatoprotective effects of hot aqueous L. africana (HALA) leaf extract (0.1-1 g/kg) and silymarin (50 mg/kg) were assessed in a model of oxidative liver damage induced by RIF and INH (100 mg/kg each) in Wistar rats for 28 days. Biochemical markers of hepatic damage such as alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were assessed. The antioxidant statuses of plasma glutathione peroxidase (GSPx), glutathione reductase (GSH), catalase (CAT) and superoxide dismutase (SOD) and lipid peroxidation were evaluated., Results: The pretreatment of INH and RIF decreased hematological indices and the antioxidant levels (P < 0.001) and increased the levels of liver marker enzymes (P < 0.001). However, pretreatment with HALA extract and silymarin provoked significant elevation of hematological indices. The levels of AST, ALT, and ALP were depressed (P < 0.001). Total triglycerides, total cholesterol, total bilirubin and low-density lipoprotein were decreased (P < 0.001). However, high-density lipoprotein, bicarbonate, and electrolytes like chloride and potassium were elevated (P < 0.001), but sodium was depressed (P < 0.05). Additionally, GSH, GSPx, SOD and CAT were elevated (P < 0.01) and malondialdehyde was depressed (P < 0.001) when compared to the RIF-INH-treated rats. Histopathological evaluations support hepatoprotective activity., Conclusion: This study demonstrated that HALA leaf extract attenuated RIF-INH-induced hepatotoxicity. L. africana could be exploited in management of RIF-INH-induced hepatitis., (Copyright © 2018 Shanghai Changhai Hospital. Published by Elsevier B.V. All rights reserved.)