Initial Event Research Articles

Biomimetic Proteoglycans Strengthen the Pericellular Matrix of Normal and Osteoarthritic Human Cartilage.

In osteoarthritis (OA), degradation of cartilage pericellular matrix (PCM), the proteoglycan-rich immediate cell microniche, is a leading event of disease initiation. This study demonstrated that biomimetic proteoglycans (BPGs) can diffuse into human cartilage from both normal and osteoarthritic donors and are preferentially localized within the PCM. Applying immunofluorescence (IF)-guided AFM nanomechanical mapping, we show that this localization of BPGs increases the PCM micromodulus of both normal and OA specimens. These results illustrate the capability of BPGs to integrate with degenerative tissues and support the translational potential of BPGs for treating human OA and other diseases associated with proteoglycan degradation.

Effects of rolling process and microstructure on the brittle crack initiation and arrest of a marine steel under low‐temperature

AbstractThe effects of the rolling process and microstructure on the brittle crack initiation and arrest of marine steel were systematically investigated. The microstructures of two steel plates produced by different rolling processes were observed and analyzed. The results demonstrated that the rolling process significantly influenced the microstructure. As the rolling temperature decreased, the grain size decreased, the high‐angle grain boundary (HAGB) content increased, and the average grain misorientation angle increased. The brittle crack initiation and the arrest abilities were evaluated by initiation fracture toughness and crack arrest toughness, respectively. The initiation fracture toughness and crack arrest toughness increased with the decreasing grain size and the increasing content of HAGBs. Finally, the influence of microstructure on the brittle crack initiation and arrest ability was explained from two aspects: (1) the critical event of crack initiation at low temperatures, and (2) the hindering effect of HAGB on the propagation of brittle cracks.

Strongly Correlated Detections of Independent Photons Eliminate Entangled Photons

Published experimental results and analytic developments indicate the possibility of achieving stronger correlations of physical events with independent photons rather than with entangled ones. The probability of coincident detections of quantum events should not be confused with the correlation of mixed states. The theoretical requirements for implementing the quantum nonlocality theory are not present in the experimental configurations purporting to prove Bohr’s or Bell’s nonlocality because of the quantum Rayleigh scattering of single photons. By means of a normalization factor corresponding to the total number of initiated events, the detection probabilities obtained experimentally are too small to enable any violation of a Bell inequality. Correlations between independent states of qubits can easily outperform those calculated with entangled photons. Additionally, the quantum joint probability for a Bell state can be factorized enabling a local detection of the alleged quantum nonlocality, if it existed

Risk-Informing Nuclear Reactor Safety: The Prediction of the Probability of Core Damage Due to Loss of Power and Cooling

Abstract The modern idea of risk-informed decision-making (RIDM) is here critically examined for all existing, advanced and, generation-IV nuclear reactor systems. Motivated by the predictive difficulties of probabilistic risk assessment (PRA) in regard to occurred accidents, it is evident that the real (not hypothetical) consequences of nuclear core damage accidents that must be considered and quantified are the financial losses, infrastructure damages, societal disruptions, and adverse political policies, and not solely the traditional exceedance of regulated radiation release or public exposure limits. With this perspective, a new dynamic analysis is presented for estimating the probability of core damage due to extended loss of power and cooling in a modern nuclear reactor, giving results different from current standardized PRA/PSA analyses. Using existing data, we approach the multitude of different events in a new way: whatever the initial event in the finite event set {external flood, fire, hurricane, ice storm, typhoon, earthquake, cyber attack…}, the fundamental concerns are the consequent loss and nonrestoration of power, and the inadequate cooling of the core. The present proposed quantitative evaluation uses applicable and fully “exchangeable” severe event data for nuclear and nonnuclear systems, including active and passive emergency back-up systems for a wide range of power losses that lead to widespread damage and societal disruption. While not design-specific, this new independent “order-of-magnitude” estimate for the probability of core damage is some two to two hundred times larger than that shown or reported in recent modern and formal PSA/PRA for licensing submissions using generic failure rate data rather than actual severe event data. This new finding suggests greater uncertainties exist than presently assumed for risk-informed decision-making (RIDM), and points to the need for a major reconsideration and updating of risk assessment and regulatory risk-informed approaches for nuclear plant safety evaluation.

Assessing risk of recurrent small bowel obstruction after non-operative management in patients with history of intra-abdominal surgery: a population-based comprehensive analysis in Taiwan.

Despite a significant 30% ten-year readmission rate for SBO patients, investigations into recurrent risk factors after non-operative management are scarce. The study aims to generate a risk factor scoring system, the 'Small Bowel Obstruction Recurrence Score' (SBORS), predicting 6-month recurrence of small bowel obstruction (SBO) after successful non-surgical management in patients who have history of intra-abdominal surgery. We analyzed data from patients aged ≥ 18 with a history of intra-abdominal surgery and diagnosed with SBO (ICD-9 code: 560, 568) and were successful treated non-surgically between 2004 and 2008. Participants were divided into model-derivation (80%) and validation (20%) group. We analyzed 23,901 patients and developed the SBORS based on factors including the length of hospital stay > 4 days, previous operations > once, hemiplegia, extra-abdominal and intra-abdominal malignancy, esophagogastric surgery and intestino-colonic surgery. Scores > 2 indicated higher rates and risks of recurrence within 6 months (12.96% vs. 7.27%, OR 1.898, p < 0.001 in model-derivation group, 12.60% vs. 7.05%, OR 1.901, p < 0.001 in validation group) with a significantly increased risk of mortality and operative events for recurrent episodes. The SBORS model demonstrated good calibration and acceptable discrimination, with an area under curve values of 0.607 and 0.599 for the score generation and validation group, respectively. We established the effective 'SBORS' to predict 6-month SBO recurrence risk in patients who have history of intra-abdominal surgery and have been successfully managed non-surgically for the initial obstruction event. Those with scores > 2 face higher recurrence rates and operative risks after successful non-surgical management.

Tuning Interactions to Control Molecular Down Conversion in [2.2]Paracyclophane Bridged Oligo‐Tetracenes

AbstractIn tetracene, the energies of the lowest singlet excited state and twice that of the lowest triplet excited state are similar, allowing both down‐conversion (i.e., singlet fission, SF) and up‐conversion (i.e., triplet‐triplet annihilation up‐conversion, TTA‐UC) processes. Through‐space and through‐bond contributions to the inter‐tetracene coupling in purposefully designed oligomers play a crucial role in determining which of the two processes dominates. In this work, the focus is exclusively on SF in newly synthesized oligo‐tetracenes linked by conjugated [2.2]paracyclophane (PCP) building blocks. By choosing different PCP substitution patterns and by varying the degree of substitution the inter‐tetracene couplings are addressed. An independent variable is connecting the tetracences to the PCP at different positions to alter the through‐bond and through‐space coupling of the resulting oligo‐tetracenes. The novel oligo‐tetracenes are investigated by means of steady‐state and time‐resolved absorption and fluorescence spectroscopies with respect to the initial events of SF, that is, the transformation of a singlet excited state into a correlated triplet pair state. Briefly, through‐space couplings are profoundly weaker than through‐bond couplings that enable the correlated triplet pair state formation. If interactions are through‐space, correlated triplet pair state formation is turned off, while it is turned on if through‐bond interactions are operative.

Don’t Call It a Failure: Systemic Risk Governance for Complex Financial Systems

The probability that an event will avalanche into an impairment of essential services constitutes a “systemic risk.” Owing to the inherent complexities of modern societies, the outbreak of a novel disease or the failure of a financial institution can rapidly escalate into an impact significantly larger than the initial event. Through the lens of complex system theory, this article draws a parallel between financial crises and disasters to contend that the regulatory framework for financial systemic risk is unequipped to address its fundamental dynamics. Epitomized by the market failure rationale, financial regulation is premised on a reductionist view that purports both systemic risk and law as external to the actions of market participants. Conversely, this article advances a twofold conceptual framework. First, it shows that systemic risk emerges from the same complex dynamics that generate the financial system. Second, it understands law as an agent of complexity, thus contributing to the emergence of finance and its inherent instability. Normatively, this conceptual framework reveals the limits of current regulatory approaches and constructs a holistic risk governance framework that is akin to the one adopted to govern disaster risks.

Spatiotemporally Mapped Endothelial Dysfunction at Bifurcations in a Coronary Artery‐on‐a‐Chip

AbstractAtherosclerotic plaques are commonly observed at low shear regions, in particular, the left main coronary artery (LMCA) bifurcation. Low shear regions at bifurcations promote endothelial dysfunction, a key factor initiating atherogenesis, however, mechanisms underlying this process are poorly understood. Dynamic in vitro models are critical to investigate endothelial dysfunction, but current static and vessels‐on‐chip systems typically lack physiologically complex geometries and local shear changes. Here, a bifurcating coronary artery‐on‐a‐chip is developed, mimicking the human LMCA, displaying reduced shear near the bifurcation, verified using computational fluid dynamics simulations. Over 7 days of dynamic culture, human coronary artery endothelial cells aligned with the flow and expressed more Endothelial nitric oxide synthase (eNOS) and intercellular cell adheison molecule‐1 (ICAM‐1) at high shear regions (12.7 dyn cm−2) adjacent to, but not at the bifurcation (0–3 dyn cm−2). After tumor necrosis factor‐alpha (TNFα) stimulation to induce endothelial dysfunction, spatially mapping cellular changes and shear gradients revealed cell alignment is disrupted over a larger area surrounding the bifurcation at a higher shear, and ICAM‐1 expression is increased closer to the bifurcation at a lower shear. This coronary artery‐on‐a‐chip establishes a system to spatially map endothelial cell behavior in response to differential shear and vessel geometry, enabling future studies into plaque initiation events, treatment targets, and drug screening.

Current Clinical Concepts: Nonoperative Management of Shoulder Instability.

Non-operative management following a shoulder dislocation or subluxation remains a challenging and complex task. Accurate diagnosis of the condition, and shared decision-making regarding operative and non-operative management, as well as timing of return to play is required. This clinical concept paper introduces a shoulder instability framework that addresses these fundamental clinical dilemmas. Valid clinical prognostic tools which can predict recurrent shoulder instability are reviewed. The process of shared decision-making within the realm of shoulder instability is also presented. Finally, a framework for progressive rehabilitation that addresses deficits in motor control, strength, and endurance in scapula and shoulder musculature is presented to guide patients from an initial instability event, through to return to play.

Firing Alterations of Neurons in Alzheimer's Disease: Are They Merely a Consequence of Pathogenesis or a Pivotal Component of Disease Progression?

Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder, yet its underlying causes remain elusive. The conventional perspective on disease pathogenesis attributes alterations in neuronal excitability to molecular changes resulting in synaptic dysfunction. Early hyperexcitability is succeeded by a progressive cessation of electrical activity in neurons, with amyloid beta (Aβ) oligomers and tau protein hyperphosphorylation identified as the initial events leading to hyperactivity. In addition to these key proteins, voltage-gated sodium and potassium channels play a decisive role in the altered electrical properties of neurons in AD. Impaired synaptic function and reduced neuronal plasticity contribute to a vicious cycle, resulting in a reduction in the number of synapses and synaptic proteins, impacting their transportation inside the neuron. An understanding of these neurophysiological alterations, combined with abnormalities in the morphology of brain cells, emerges as a crucial avenue for new treatment investigations. This review aims to delve into the detailed exploration of electrical neuronal alterations observed in different AD models affecting single neurons and neuronal networks.

Do the same chlorinated organophosphorus flame retardants that cause cytotoxicity and DNA damage share the same pathway?

Organophosphorus flame retardants (OPFRs) have been frequently detected with relatively high concentrations in various environmental media and are considered emerging environmental pollutants. However, their biological effect and underlying mechanism is still unclear, and whether chlorinated OPFRs (Cl-OPFRs) cause adverse outcomes with the same molecular initial events or share the same key events (KEs) remains unknown. In this study, in vitro bioassays were conducted to analyze the cytotoxicity, mitochondrial impairment, DNA damage and molecular mechanisms of two Cl-OPFRs. The results showed that these two Cl-OPFRs, which have similar structures, induced severe cellular and molecular damages via different underlying mechanisms. Both tris(2-chloroethyl) phosphate (TCEP) and tris(1-chloro-2-propyl) (TCPP) induced oxidative stress-mediated mitochondrial impairment and DNA damage, as shown by the overproduction of intracellular reactive oxygen species (ROS) and mitochondrial superoxide. Furthermore, the DNA damage caused by TCPP resulted in p53/p21-mediated cell cycle arrest, as evidenced by flow cytometry and real-time PCR. At the cellular and molecular levels, TCPP increased the sub-G1 apoptotic peak and upregulated the p53/Bax apoptosis pathway, possibly resulted in apoptosis associated with its stronger cytotoxicity. Although structurally similar to TCPP, TCEP did not induce mitochondrial impairment and DNA damage by the same KEs. These results provide insight into the toxicity of Cl-OPFRs with similar structures but different mechanisms, which is of great significance for constructing adverse outcome pathways or determining intermediate KEs.

Is Gillies Temporal Incision Necessary to Successfully Reduce Zygomaticomaxillary Fracture Combined With Orbital Wall Fracture?

Gillies temporal approach has been widely used to reduce zygoma fractures. Traditionally, it is the preferred method to reduce the isolated zygomatic arch fracture. It can be combined with other approaches, such as subciliary and intraoral incisions for more complex zygomatic fractures, so-called tripod fractures. In this study, the author hypothesized that complex zygomaticomaxillary bone fracture reduction can successfully be achieved without Gillies temporal incision. The author retrospectively analyzed the medical records and 3D facial Computed Tomography (CT) scans of 80 patients who had orbitozygomaticomaxillary fractures that affected >3 sites among the frontozygomatic suture, zygomaticomaxillary buttress, zygomatic arch, and orbital walls from May 2021 to August 2023. A single surgeon performed all surgical operations. All fractures were reduced with 2 incisions, lower eyelid incisions (subciliary or subconjunctival) and intraoral incisions (gingivobuccal approach), within a week of the initial traumatic event. After sufficient exposure to fractured sites using 2 incisions, the author reduced the fracture with a bony hook at the inferior orbital rim, lateral wall, or Boise elevator at the intraoral incision. All cases were successfully corrected without any complications. In conclusion, the author can successfully reduce complex zygomaticomaxillary fracture combined with orbital wall fractures without temporal Gillies approach.

Comparative proteomic analysis of seed germination between allotetraploid cotton Gossypium hirsutum and Gossypium barbadense

Seed germination, a key initial event in the plant life cycle, directly affects cotton yield and quality. Gossypium barbadense and Gossypium hirsutum gradually evolved through polyploidization, resulting in different characteristics, and this interspecific variation lacks genetic and molecular explanation. This work aimed to compare the proteomes between G. barbadense and G. hirsutum during seed germination. Here, we identified 2740 proteins for G. barbadense and 3758 for G. hirsutum. In the initial state, proteins in two cotton involved similar bioprocess, such as sugar metabolism, DNA repairing, and ABA signaling pathway. However, in the post-germination stage, G. hirsutum expressed more protein related to redox homeostasis, peroxidase activity, and pathogen interactions. Analyzing the different expression patterns of 915 single-copy orthogroups between the two kinds of cotton indicated that most of the differentially expressed proteins in G. barbadense were related to carbon metabolism. In contrast, most proteins in G. hirsutum were associated with stress response. Besides that, by proteogenomic analysis, we found 349 putative non-canonical peptides, which may be involved in plant development. These results will help to understand the different characteristics of these two kinds of cotton, such as fiber quality, yield, and adaptability. Significance statementCotton is the predominant natural fiber crop worldwide; Gossypium barbadense and Gossypium hirsutum have evolved through polyploidization to produce differing traits. However, given their specific features, the divergence of mechanisms underlying seed germination between G. hirsutum and G. barbadense has not been discussed. Here, we explore what protein contributes to interspecific differences between G. barbadense and G. hirsutum during the seed germination period. This study helps to elucidate the evolution and domestication history of cotton polyploids and may allow breeders to understand their domestication history better and improve fiber quality and adaptability.

A Practical Approach to Automatic Earthquake Catalog Compilation in Local OBS Networks Using Deep-Learning and Network-Based Algorithms

Abstract In land-based seismology, modern automatic earthquake detection and phase picking algorithms have already proven to outperform classic approaches, resulting in more complete catalogs when only taking a fraction of the time needed for classic methods. For marine-based seismology, similar advances have not been made yet. For ocean-bottom seismometer (OBS) data, additional challenges arise, such as a lower signal-to-noise ratio and fewer labeled data sets available for training deep-learning models. However, the performance of available deep-learning models has not yet been extensively tested on marine-based data sets. Here, we apply three different modern event detection and phase picking approaches to an ∼12 month local OBS data set and compare the resulting earthquake catalogs and location results. In addition, we evaluate their performance by comparing different subcatalogs of manually detected events and visually revised picks to their automatic counterparts. The results show that seismicity patterns from automatically compiled catalogs are comparable to a manually revised catalog after applying strict location quality control criteria. However, the number of such well-constrained events varies between the approaches and catalog completeness cannot be reliably determined. We find that PhaseNet is more suitable for local OBS networks compared with EQTransformer and propose a pick-independent event detection approach, such as Lassie, as the preferred choice for an initial event catalog compilation. Depending on the aim of the study, different schemes of manual repicking should be applied because the automatic picks are not yet reliable enough for developing a velocity model or interpreting small-scale seismicity patterns.

Risk factors for poor outcome after aneurysmal subarachnoid hemorrhage in patients with initial favorable neurological status

BackgroundAneurysmal subarachnoid hemorrhage (aSAH) remains a devastating diagnosis. A poor outcome is known to be highly dependent on the initial neurological status. Our goal was to identify other parameters that favor the risk of complications and poor outcome in patients with aSAH and initially favorable neurologic status.MethodsConsecutive aSAH cases treated at our hospital between 01/2003 and 06/2016 with the initial World Federation of Neurosurgical Societies grades I–III were included. Data on demographic characteristics, previous medical history, initial aSAH severity, and functional outcome after aSAH were collected. The study endpoints were the occurrence of cerebral infarcts, in-hospital mortality, and unfavorable outcome at 6 months after aSAH (modified Rankin scale > 3).ResultsIn the final cohort (n= 582), the rate of cerebral infarction, in-hospital mortality, and unfavorable outcome was 35.1%, 8.1%, and 17.6% respectively. The risk of cerebral infarction was independently related to the presence of acute hydrocephalus (adjusted odds ratio [aOR]=2.33, p<0.0001), aneurysm clipping (aOR=1.78, p=0.003), and use of calcium channel blockers concomitant to nimodipine (aOR=2.63, p=0.002). Patients’ age (>55 years, aOR=4.24, p<0.0001), acute hydrocephalus (aOR=2.43, p=0.036), and clipping (aOR=2.86, p=0.001) predicted in-hospital mortality. Baseline characteristics associated with unfavorable outcome at 6 months were age (aOR=2.77, p=<0.0001), Fisher grades III–IV (aOR=2.81, p=0.016), acute hydrocephalus (aOR=2.22, p=0.012), clipping (aOR=3.98, p<0.0001), admission C-reactive protein>1mg/dL (aOR=1.76, p=0.035), and treatment intervals (aOR=0.64 per-5-year-intervals, p=0.006).ConclusionsAlthough cerebral infarction is a common complication in aSAH individuals with favorable initial clinical condition, >80% of these patients show favorable long-term outcome. The knowledge of outcome-relevant baseline characteristics might help to reduce the burden of further complications and poor outcome in aSAH patients who tolerated the initial bleeding event well.

Nontargeted Identification of Organic Components in Fine Particulate Matter Related to Lung Tumor Metastasis Based on an Adverse Outcome Pathway Strategy.

Emerging studies implicate fine particulate matter (PM2.5) and its organic components (OCs) as urgent hazard factors for lung cancer progression in nonsmokers. Establishing the adverse outcome pathway (AOP)-directed nontargeted identification method, this study aimed to explore whether PM2.5 exposure in coal-burning areas promoted lung tumor metastasis and how we identify its effective OCs to support traceability and control of regional PM2.5 pollution. First, we used a nude mouse model of lung cancer for PM2.5 exposure and found that the exposure significantly promoted the hematogenous metastases of A549-Luc cells in lung tissues and the adverse outcomes (AOs), with key events (KEs) including the changed expression of epithelial-mesenchymal transition (EMT) markers, such as suppression of E-cad and increased expression of Fib. Subsequently, using AOs and KEs as adverse outcome directors, we identified a total of 35 candidate chemicals based on the in vitro model and nontargeted analysis. Among them, tributyl phosphate (C12H27O4P), 2-bromotetradecane (C14H29Br), and methyl decanoate (C11H22O2) made greater contributions to the AOs. Finally, we clarified the interactions between these OCs and EMT-activating transcription factors (EMT-ATFs) as the molecular initiation event (MIE) to support the feasibility of the above identification strategy. The present study updates a new framework for identifying tumor metastasis-promoting OCs in PM2.5 and provides solid data for screening out chemicals that need priority control in polluted areas posing higher lung cancer risk.

Longitudinal Assessment of Hair Cortisol as a Predictor of Psychological Symptoms During COVID-19

BackgroundThere is a lack of evidence regarding enduring psychoneuroendocrine changes following an initial traumatic event, particular in the presence of an ongoing stressor. The coronavirus pandemic presents an opportunity to explore this matter. Consequently, the purpose of the present study was to investigate the impact of the ongoing pandemic (2021) on individuals, who experienced a first-time motor vehicle crash (MVC) at least 6 years earlier. To this end, we hypothesized that hair cortisol concentrations (HCC) following a first-time traumatic event positively predict symptoms of depression. MethodWe investigated N = 69 individuals (18 – 65 yrs.), who were victims of a MVC during 2010 – 2014. Hair strands were collected 10 days (t1) and 3 months after the MVC (t2), as well during the pandemic in 2021 (t3). To assess symptoms of depression, the participants filled out the Beck Depression Inventory at t1 - t3 and were additionally interviewed (Structured Clinical Interview for DSM-IV Axis I) at t1 and t2. Exclusion criteria conveyed a lifetime or acute mental disorder (incl. past trauma exposure). ResultsElevated pre-pandemic HCC following adversity (i.e., MVC) significantly predicted symptoms of depression in adults during the coronavirus pandemic (BDI: ß =.44, p =.010, R2 =.20), even after controlling for confounders. HCC significantly decreased over time, while in average psychological symptoms remained consistent. ConclusionCortisol dysregulation in the past presents an enduring vulnerability to ongoing stress. In this regard, vulnerable groups may benefit from preventive measures. This finding validates the predictive power of HCC and extended past evidence in this regard, at the same time reinforcing the concept of the diathesis-stress model.

Time-resolved role of P2X4 and P2X7 during CD8+ T cell activation.

CD8+ T cells are a crucial part of the adaptive immune system, responsible for combating intracellular pathogens and tumor cells. The initial activation of T cells involves the formation of highly dynamic Ca2+ microdomains. Recently, purinergic signaling was shown to be involved in the formation of the initial Ca2+ microdomains in CD4+ T cells. In this study, the role of purinergic cation channels, particularly P2X4 and P2X7, in CD8+ T cell signaling from initial events to downstream responses was investigated, focusing on various aspects of T cell activation, including Ca2+ microdomains, global Ca2+ responses, NFAT-1 translocation, cytokine expression, and proliferation. While Ca2+ microdomain formation was significantly reduced in the first milliseconds to seconds in CD8+ T cells lacking P2X4 and P2X7 channels, global Ca2+ responses over minutes were comparable between wild-type (WT) and knockout cells. However, the onset velocity was reduced in P2X4-deficient cells, and P2X4, as well as P2X7-deficient cells, exhibited a delayed response to reach a certain level of free cytosolic Ca2+ concentration ([Ca2+]i). NFAT-1 translocation, a crucial transcription factor in T cell activation, was also impaired in CD8+ T cells lacking P2X4 and P2X7. In addition, the expression of IFN-γ, a major pro-inflammatory cytokine produced by activated CD8+ T cells, and Nur77, a negative regulator of T cell activation, was significantly reduced 18h post-stimulation in the knockout cells. In line, the proliferation of T cells after 3 days was also impaired in the absence of P2X4 and P2X7 channels. In summary, the study demonstrates that purinergic signaling through P2X4 and P2X7 enhances initial Ca2+ events during CD8+ T cell activation and plays a crucial role in regulating downstream responses, including NFAT-1 translocation, cytokine expression, and proliferation on multiple timescales. These findings suggest that targeting purinergic signaling pathways may offer potential therapeutic interventions.

Human Adenovirus Entry and Early Events during Infection of Primary Murine Neurons: Immunofluorescence Studies In Vitro.

Human adenovirus (HAdV) is a common pathogen, which can lead to various clinical symptoms and-in some cases-central nervous system (CNS) dysfunctions, such as encephalitis and meningitis. Although the initial events of virus entry have already been identified in various cell types, the mechanism of neuronal uptake of adenoviruses is relatively little understood. The aim of this study was to investigate early events during adenoviral infection, in particular to determine the connection between cellular coxsackievirus and adenovirus receptor (CAR), clathrin, caveolin, and early endosomal proteins (EEA1 and Rab5) with the entry of HAdVs into primary murine neurons in vitro. An immunofluorescence assay and confocal microscopy analysis were carried out to determine HAdV4, 5, and 7 correlation with CAR, clathrin, caveolin, and early endosomal proteins in neurons. The quantification of Pearson's coefficient between CAR and HAdVs indicated that the HAdV4 and HAdV5 types correlated with CAR and that the correlation was more substantial for HAdV5. Inhibition of clathrin-mediated endocytosis using chlorpromazine limited the infection with HAdV, whereas inhibition of caveolin-mediated endocytosis did not affect virus entry. Thus, the entry of tested HAdV types into neurons was most likely associated with clathrin but not caveolin. It was also demonstrated that HAdVs correlate with the Rab proteins (EEA1, Rab5) present in early vesicles, and the observed differences in the manner of correlation depended on the serotype of the virus. With our research, we strove to expand knowledge regarding the mechanism of HAdV entry into neurons, which may be beneficial for developing potential therapeutics in the future.

Diversification and dispersal in the Americas revealed by new phylogenies of the wrens and allies (Passeriformes: Certhioidea)

Abstract The passerine superfamily Certhioidea lacks a complete phylogeny despite decades of recognition as a clade and extensive systematic work within all its constituent families. Here, we inferred a near-complete species-level phylogeny of Certhioidea from a molecular supermatrix, including the first comprehensive sampling of the wrens (Troglodytidae), and used this phylogeny to infer its biogeographic and diversification histories. We also inferred an expanded phylogeny including nearly 100 putative phylospecies previously documented in the literature, and we found that including this diversity had notable impacts on the inferred evolutionary history of Certhioidea. This phylospecies-level tree documented a few instances of species paraphyly, some previously described in the literature and some novel. We found that Certhioidea originated largely in Eurasia and dispersed into North America five times in the last 20 million years, including at the origin of the “New World certhioids,” wrens and gnatcatchers, a clade herein named Orthourae. After this initial dispersal event, both wrens and gnatcatchers diversified extensively across the hemisphere, with both lineages repeatedly crossing between continents. However, we detected no notable impact of the formation of the Isthmus of Panama on the frequency of dispersal events between North and South America. The inclusion of phylospecies altered this biogeographic inference in some portions of the tree but overall was largely consistent. With species-level sampling, we found that diversification rates within Certhioidea were largely constant through time with a detectable deceleration toward the present. By contrast, phylospecies-level sampling recovered a different diversification history with a significant rate increase at the crown node of Orthourae after dispersing into the Americas and increased speciation rates particularly within the genera Polioptila and Henicorhina. This largely resolved phylogeny for Certhioidea has yielded important insights into the evolutionary history of this group and provides a framework for future comparative work on this fascinating clade.