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Because the COVID-19 pandemic has been implicated in increased mental health concerns for families of low income, we aimed to describe maternal perspectives about the pandemic's impact on their kindergartener's mental health during the 2020-2021 school year. We conducted 22 in-depth, semi-structured interviews with U.S. mothers with low income who had kindergarten-age children (50% male and 50% female). All participants were female, ranging in age from 24 to 44 years, and reported the following ethnic/racial identities: non-Hispanic Black or African American (41%), Hispanic of any race (36%) and non-Hispanic, White (23%). With a team comprising multiple researchers from varied disciplines (e.g., medicine, education and public health) our analytic process used an iterative approach for developing and revising codes and themes until we reached thematic saturation. Most mothers described negative impacts on social, behavioural and emotional aspects of mental health. Some described positive social impacts, including strengthened family relationships. Mothers described no positive changes to behavioural or emotional aspects of mental health. Maternal perspectives suggest the importance of prioritising access to screening and treating mental health needs to support children's kindergarten transition, mitigate pandemic impacts and plan for future disruptions.

BackgroundRecently shown to regulate cardiac development, the secreted axon guidance molecule SLIT3 maintains its expression in the postnatal heart. Despite its known expression in the cardiovascular system after birth, SLIT3's relevance to cardiovascular function in the postnatal state remains unknown. As such, the objectives of this study were to determine the postnatal myocardial sources of SLIT3 and to evaluate its functional role in regulating the cardiac response to pressure overload stress.MethodsWe performed in vitro studies on cardiomyocytes and myocardial tissue samples from patients and performed in vivo investigation with SLIT3 and ROBO1 (roundabout homolog 1) mutant mice undergoing transverse aortic constriction to establish the role of SLIT3-ROBO1 in adverse cardiac remodeling.ResultsWe first found that SLIT3 transcription was increased in myocardial tissue obtained from patients with congenital heart defects that caused ventricular pressure overload. Immunostaining of hearts from WT (wild-type) and reporter mice revealed that SLIT3 is secreted by cardiac stromal cells, namely fibroblasts and vascular mural cells, within the heart. Conditioned media from cardiac fibroblasts and vascular mural cells both stimulated cardiomyocyte hypertrophy in vitro, an effect that was partially inhibited by an anti-SLIT3 antibody. Also, the N-terminal, but not the C-terminal, fragment of SLIT3 and the forced overexpression of SLIT3 stimulated cardiomyocyte hypertrophy and the transcription of hypertrophy-related genes. We next determined that ROBO1 was the most highly expressed roundabout receptor in cardiomyocytes and that ROBO1 mediated SLIT3's hypertrophic effects in vitro. In vivo, Tcf21+ fibroblast and Tbx18+ vascular mural cell-specific knockout of SLIT3 in mice resulted in decreased left ventricular hypertrophy and cardiac fibrosis after transverse aortic constriction. Furthermore, α-MHC+ cardiomyocyte-specific deletion of ROBO1 also preserved left ventricular function and abrogated hypertrophy, but not fibrosis, after transverse aortic constriction.ConclusionsCollectively, these results indicate a novel role for the SLIT3-ROBO1-signaling axis in regulating postnatal cardiomyocyte hypertrophy induced by pressure overload.

Marta Kutwin,1 Malwina Sosnowska-Ławnicka,1 Barbara Nasiłowska,2 Agata Lange,1 Mateusz Wierzbicki,1 Sławomir Jaworski1 1Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland; 2Institute of Optoelectronics, Military University of Technology, Warsaw, PolandCorrespondence: Marta Kutwin, Email marta_kutwin@sggw.edu.plPurpose: The use of nanotechnology in medicine has gained attention in developing drug delivery systems. GO has the potential to deliver microRNA (miRNA) mimics or antisense structures. MiRNAs regulate gene expression and their dysregulation is implicated in diseases, including cancer. This study aims to observe changes in morphology, viability, mRNA expression of mTOR/PI3K/Akt and PTEN genes in U87, U118, U251, A172 and T98 glioblastoma cells and xenograft models after GO self-assembly with mimic miRNA-7.Methods: Colloidal suspension of graphene oxide (GO) was used for obtaining the GO-mimic miRNA-7 nanosystems by self-assembly method. The ultrastructure, size distribution and ATR-FTIR and UV-Vis spectrum were analyzed. The Zeta potential was measured to verify the stability of obtained nanosystem. The entrapment efficiency, loading capacity and released kinetics of mimic miRNA-7 form GO-mimic miRNA-7 nanosystems were analyzed. The transfection efficiency into the glioblastoma cell lines U87, U118, U251, A172 and T98 of mimic miRNA-7 delivered by GO nanosystems was measure by confocal microscopy and flow cytometry. The changes at mRNA expression level of mTOR, PI3K, AKT1 and PTEN genes was measured by qPCR analysis. The xenograft model of U87 and A172 tumour tissue was performed to analyze the effect at tumor size and volume after GO- mimic miRNA-7 nanosystem administration.Results: The ultrastructure of GO-mimic miRNA-7 nanosystems showed high affinity of mimic miRNA into the GO. The results of transfection efficiency, cell morphology and viability showed that GO -miRNA-7 effectively deliver mimics miRNA-7 into U87, U118, U251, A172 and T98 glioblastoma cells. This approach can reverse miRNA-7 expression’s downstream effects and target the mTOR PI3K/Akt pathway observed at gene expression level, reducing xenograft tumour size and volume.Conclusion: The findings of the study could have significant implications for the development of advanced and precise GO based nanosystems specifically designed for miRNA therapy in cancer treatment.Keywords: GO, micro-RNA, cancer, mTOR/PI3K/Akt

Joanna Cymerys,1,* Michalina Bartak,1,* Anna Słońska,1 Agata Lange,2 Sławomir Jaworski,2 Marcin Chodkowski,3 Agnieszka Ostrowska,2 Mateusz Wierzbicki,2 Ewa Sawosz,2 Marcin W Bańbura1 1Department of Preclinical Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland; 2Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland; 3Military Institute of Hygiene and Epidemiology, Warsaw, Poland*These authors contributed equally to this workCorrespondence: Joanna Cymerys, Department of Preclinical Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences, 02-786, Warsaw, Poland, Email joanna_cymerys@sggw.edu.plBackground: The high infectivity of coronaviruses has led to increased interest in developing new strategies to prevent virus spread. Silver nanoparticles (AgNPs) and graphene oxide (GO) have attracted much attention in the antiviral field. We investigated the potential antiviral activity of GO and AgNPs combined in the nanocomposite GO-Ag against murine betacoronavirus MHV using an in vitro model.Methods: GO, AgNPs, and GO-Ag characterization (size distribution, zeta potential, TEM visualization, FT-IR, and EDX analysis) and XTT assay were performed. The antiviral activity of GO-Ag nanocomposites was evaluated by RT-qPCR and TCID50 assays. The results were compared with free AgNPs and pure GO. Cell growth and morphology of MHV-infected hepatocytes treated with GO-Ag composites were analyzed by JuLI™Br. Immunofluorescence was used to visualize the cell receptor used by MHV. Ultrastructural SEM analysis was performed to examine cell morphology after MHV infection and GO-Ag composite treatment.Results: A significant reduction in virus titer was observed for all nanocomposites tested, ranging from 3.2 to 7.3 log10 TCID50. The highest titer reduction was obtained for GO 5 μg/mL - Ag 25 μg/mL in the post-treatment method. These results were confirmed by RT-qPCR analysis. The results indicate that GO-Ag nanocomposites exhibited better antiviral activity compared to AgNPs and GO. Moreover, the attachment of AgNPs to the GO flake platform reduced their cytotoxicity. In addition, the GO-Ag composite modulates the distribution of the Ceacam1 cell receptor and can modulate cell morphology.Conclusion: Graphene oxide sheets act as a stabilizing agent, inhibiting the accumulation of AgNPs and reducing their cellular toxicity. The GO-Ag composite can physically bind and inhibit murine betacoronavirus from entering cells. Furthermore, the constant presence of GO-Ag can inhibit MHV replication and significantly limit its extracellular release. In conclusion, GO-Ag shows promise as an antiviral coating on solid surfaces to minimize virus transmission and spread. Keywords: graphene oxide, silver nanoparticles, GO-Ag composite, antiviral, coronavirus, MHV

Abstract Background Mosquito host feeding patterns are an important factor of the species-specific vector capacity determining pathogen transmission routes. Culex pipiens s.s./Cx. torrentium are competent vectors of several arboviruses, such as West Nile virus and Usutu virus. However, studies on host feeding patterns rarely differentiate the morphologically indistinguishable females. Methods We analyzed the host feeding attraction of Cx. pipiens and Cx. torrentium in host-choice studies for bird, mouse, and a human lure. In addition, we summarized published and unpublished data on host feeding patterns of field-collected specimens from Germany, Iran, and Moldova from 2012 to 2022, genetically identified as Cx. pipiens biotype pipiens, Cx. pipiens biotype molestus, Cx. pipiens hybrid biotype pipiens × molestus, and Cx. torrentium, and finally put the data in context with similar data found in a systematic literature search. Results In the host-choice experiments, we did not find a significant attraction to bird, mouse, and human lure for Cx. pipiens pipiens and Cx. torrentium. Hosts of 992 field-collected specimens were identified for Germany, Iran, and Moldova, with the majority determined as Cx. pipiens pipiens, increasing the data available from studies known from the literature by two-thirds. All four Culex pipiens s.s./Cx. torrentium taxa had fed with significant proportions on birds, humans, and nonhuman mammals. Merged with the data from the literature from 23 different studies showing a high prevalence of blood meals from birds, more than 50% of the blood meals of Cx. pipiens s.s. were identified as birds, while up to 39% were human and nonhuman mammalian hosts. Culex torrentium fed half on birds and half on mammals. However, there were considerable geographical differences in the host feeding patterns. Conclusions In the light of these results, the clear characterization of the Cx. pipiens s.s./Cx. torrentium taxa as ornithophilic/-phagic or mammalophilic/-phagic needs to be reconsidered. Given their broad host ranges, all four Culex taxa could potentially serve as enzootic and bridge vectors. Graphical Abstract

Abstract Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to successfully eradicate BLV through improved detection and management methods. However, with the increasing novel molecular detection methods there have been few efforts to standardize these results at global scale. This study aimed to determine the interlaboratory accuracy and agreement of 11 molecular tests in detecting BLV. Each qPCR/ddPCR method varied by target gene, primer design, DNA input and chemistries. DNA samples were extracted from blood of BLV-seropositive cattle and lyophilized to grant a better preservation during shipping to all participants around the globe. Twenty nine out of 44 samples were correctly identified by the 11 labs and all methods exhibited a diagnostic sensitivity between 74 and 100%. Agreement amongst different assays was linked to BLV copy numbers present in samples and the characteristics of each assay (i.e., BLV target sequence). Finally, the mean correlation value for all assays was within the range of strong correlation. This study highlights the importance of continuous need for standardization and harmonization amongst assays and the different participants. The results underscore the need of an international calibrator to estimate the efficiency (standard curve) of the different assays and improve quantitation accuracy. Additionally, this will inform future participants about the variability associated with emerging chemistries, methods, and technologies used to study BLV. Altogether, by improving tests performance worldwide it will positively aid in the eradication efforts.

Abstract Atrial fibrillation (A-fib) is the most common type of heart arrhythmia, typically treated with radiofrequency catheter ablation to isolate the heart from abnormal electrical signals. Monitoring the formation of ablation-induced lesions is crucial for preventing recurrences and complications arising from excessive or insufficient ablation. Existing imaging modalities lack real-time feedback, and their intraoperative usage is in its early stages. A critical need exists for an imaging-based lesion indexing (LSI) method that directly reflects tissue necrosis formation. Previous studies have indicated that spectroscopic photoacoustic (sPA) imaging can differentiate ablated tissues from their non-ablated counterparts based on PA spectrum variation. In this paper, we introduce a method for detecting ablation lesion boundaries using sPA imaging. This approach utilizes ablation LSI, which quantifies the ratio between the signal from ablated tissue and the total tissue signal. We enhance boundary detection accuracy by adapting a regression model-based compensation. Additionally, the method was cross-validated with clinically used intraoperative monitoring parameters. The proposed method was validated with ex vivo porcine cardiac tissues with necrotic lesions created by different ablation durations. The PA-measured lesion size was compared with gross pathology. Statistical analysis demonstrates a strong correlation (R > 0.90) between the PA-detected lesion size and gross pathology. The PA-detected lesion size also exhibits a moderate to strong correlation (R > 0.75) with local impedance changes recorded during procedures. These results suggest that the introduced PA imaging-based LSI has great potential to be incorporated into the clinical workflow, guiding ablation procedures intraoperatively.

Dominic Jaworski,1,2 Edward J Park1,2 1Mechatronic Systems Engineering, Simon Fraser University, Surrey, BC, V3T 0A3, Canada; 2WearTech Labs, Simon Fraser University, Surrey, BC, V3V 0C6, CanadaCorrespondence: Edward J Park; Dominic Jaworski, Surrey Centre 2 Building, 9639 - 137A Street, Unit 206, Surrey, BC, V3V 0C6, Canada, Email ed_park@sfu.ca; djaworsk@sfu.caPurpose: Wearable or non-contact, non-intrusive devices present a practical alternative to traditional polysomnography (PSG) for daily assessment of sleep quality. Physiological signals have been known to be nonlinear and nonstationary as the body adapts to states of rest or activity. By integrating more sophisticated nonlinear methodologies, the accuracy of sleep stage identification using such devices can be improved. This advancement enables individuals to monitor and adjust their sleep patterns more effectively without visiting sleep clinics.Patients and Methods: Six participants slept for three cycles of at least three hours each, wearing PSG as a reference, along with an Apple Watch, an actigraphy device, and a ballistocardiography (BCG) bed sensor. The physiological signals were processed with nonlinear methods and trained with a long short-term memory (LSTM) model to classify sleep stages. Nonlinear methods, such as return maps with advanced techniques to analyze the shape and asymmetry in physiological signals, were used to relate these signals to the autonomic nervous system (ANS). The changing dynamics of cardiac signals in restful or active states, regulated by the ANS, were associated with sleep stages and quality, which were measurable.Results: Approximately 73% agreement was obtained by comparing the combination of the BCG and Apple Watch signals against a PSG reference system to classify rapid eye movement (REM) and non-REM sleep stages.Conclusion: Utilizing nonlinear methods to evaluate cardiac dynamics showed an improved sleep quality detection with the non-intrusive devices in this study. A system of non-intrusive devices can provide a comprehensive outlook on health by regularly measuring sleeping patterns and quality over time, offering a relatively accessible method for participants. Additionally, a non-intrusive system can be integrated into a user’s or clinic’s bedroom environment to measure and evaluate sleep quality without negatively impacting sleep. Devices placed around the bedroom could measure user vitals over longer periods with minimal interaction from the user, representing their natural sleeping trends for more accurate health and sleep disorder diagnosis.Keywords: ballistocardiogram, wearable, heart rate variability, nonstationary signals

Abstract The conventional design of steel structure objects relies on a first-order elastic analysis, where the entire object is treated as a set of individual structural elements requiring time-consuming, semi-empirical design calculations. Such an approach leads to inefficient design time and excessive material consumption and may additionally result in designing on the verge of structural safety. The AEC sector's technological and digitization advancement process forces designers to use advanced design methods. Hence, it is necessary to indicate the benefits of using effective optimization. The paper presents a comparative analysis of steel domes using two design approaches: traditional first-order analysis and an advanced second-order analysis. The latter method considers the influence of structural deformation on the magnitude of internal forces. Eight models were developed, varying in terms of the connection's stiffness. The work results identify the differences between the two selected design approaches and present opportunities for further structural performance of steel structures.

In this paper, we draw on our recent research to inspect again some of the theoretical perspectives we have been using to analyze data and to characterize teaching-learning in university settings. We focus particularly, within a sociocultural perspective on Activity Theory and the construct the "Teaching Triad," seeking to embed the Triad within an Activity Theory perspective. To achieve this, we relate the Teaching Triad with aspects of the sociocultural setting both in and beyond direct interactions in face-to-face teaching. While this is mainly a theoretical paper, an example is taken from observations of teaching in university lectures in a Greek university to show how these theoretical perspectives have provided insights to the institutional and cultural complexities involved and in what ways the Triad construct has evolved.

The dorsal funiculus in the spinal cord relays somatosensory information to the brain. It is made of T-shaped bifurcation of dorsal root ganglion (DRG) sensory axons. Our previous study has shown that Slit signaling is required for proper guidance during bifurcation, but loss of Slit does not affect all DRG axons. Here, we examined the role of the extracellular molecule Netrin-1 (Ntn1). Using wholemount staining with tissue clearing, we showed that mice lacking Ntn1 had axons escaping from the dorsal funiculus at the time of bifurcation. Genetic labeling confirmed that these misprojecting axons come from DRG neurons. Single axon analysis showed that loss of Ntn1 did not affect bifurcation but rather altered turning angles. To distinguish their guidance functions, we examined mice with triple deletion of Ntn1, Slit1, and Slit2 and found a completely disorganized dorsal funiculus. Comparing mice with different genotypes using immunolabeling and single axon tracing revealed additive guidance errors, demonstrating the independent roles of Ntn1 and Slit. Moreover, the same defects were observed in embryos lacking their cognate receptors. These in vivo studies thus demonstrate the presence of multi-factorial guidance mechanisms that ensure proper formation of a common branched axonal structure during spinal cord development.

Surgeries and procedures in otolaryngology require close collaboration between anesthesiologists and ENT specialists, especially for operations involving the larynx and airways, which pose challenges for maintaining airway patency and visibility. Choosing the type of anesthesia, whether general or local, is crucial. This study examines the benefits and drawbacks of both types of anesthesia in tonsillectomy and adenoidectomy, tympanostomy and myringotomy, and flexible endoscopic biopsy of the pharynx and larynx. A review of current guidelines, systematic reviews and relevant clinical cohort studies was conducted. The findings highlight several advantages of local anesthesia, including shorter operation times, reduced hospital stays, lower costs, decreased use of analgesics, and improved patient comfort.

Thermoacoustic systems represent a promising technology for clean cooling and/or power generation. However, understanding of the behavior of oscillatory flow inside the system under high drive ratio operating conditions, at elevated mean pressures, range of frequencies, and high flow amplitudes, remains limited and poses challenges for designing efficient systems with minimal losses. This paper presents a numerical investigation into the flow and heat transfer characteristics across a coupled cold and hot parallel-plate heat exchanger operating under standing-wave type oscillatory flow conditions. The models were developed for various conditions, spanning frequencies from 13 Hz to 100 Hz and mean pressures between 1 and 20 bar, using both nitrogen and helium as working fluids. The study reveals significant phenomena such as heat accumulation and annular effects in both temperature and velocity/vorticity fields. These findings were discussed in the context of their impact on the heat transfer performance of the heat exchanger. Specifically, the emergence of vortices was examined in relation to mean pressure, alongside jet-like velocity profiles and natural convection effects. Furthermore, the paper includes a comparative analysis between the heat transfer model employed in this study and previous works, aiming to validate the findings. This research enhances the understanding of fluid dynamics and heat transfer phenomena within heat exchangers of thermoacoustic devices, thereby contributing to advancements in future design improvements.

Understanding the molecular basis of innate immune evasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an important consideration for designing the next wave of therapeutics. Here, we investigate the role of the nonstructural protein 16 (NSP16) of SARS-CoV-2 in infection and pathogenesis. NSP16, a ribonucleoside 2'-O-methyltransferase (MTase), catalyzes the transfer of a methyl group to mRNA as part of the capping process. Based on observations with other CoVs, we hypothesized that NSP16 2'-O-MTase function protects SARS-CoV-2 from cap-sensing host restriction. Therefore, we engineered SARS-CoV-2 with a mutation that disrupts a conserved residue in the active site of NSP16. We subsequently show that this mutant is attenuated both in vitro and in vivo, using a hamster model of SARS-CoV-2 infection. Mechanistically, we confirm that the NSP16 mutant is more sensitive than wild-type SARS-CoV-2 to type I interferon (IFN-I) in vitro. Furthermore, silencing IFIT1 or IFIT3, IFN-stimulated genes that sense a lack of 2'-O-methylation, partially restores fitness to the NSP16 mutant. Finally, we demonstrate that sinefungin, an MTase inhibitor that binds the catalytic site of NSP16, sensitizes wild-type SARS-CoV-2 to IFN-I treatment and attenuates viral replication. Overall, our findings highlight the importance of SARS-CoV-2 NSP16 in evading host innate immunity and suggest a target for future antiviral therapies. IMPORTANCE Similar to other coronaviruses, disruption of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) NSP16 function attenuates viral replication in a type I interferon-dependent manner. In vivo, our results show reduced disease and viral replication at late times in the hamster lung, but an earlier titer deficit for the NSP16 mutant (dNSP16) in the upper airway. In addition, our results confirm a role for IFIT1 but also demonstrate the necessity of IFIT3 in mediating dNSP16 attenuation. Finally, we show that targeting NSP16 activity with a 2'-O-methyltransferase inhibitor in combination with type I interferon offers a novel avenue for antiviral development.