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Abstract Enterovirus D68 (EV-D68) has emerged as a significant cause of acute respiratory illness in children globally, notably following its extensive outbreak in North America in 2014. A recent outbreak of EV-D68 was observed in Ontario, Canada, from August to October 2022. Our phylogenetic analysis revealed a notable genetic similarity between the Ontario outbreak and a concurrent outbreak in Maryland, USA. Utilizing Bayesian phylodynamic modeling on whole genome sequences (WGS) from both outbreaks, we determined the median peak time-varying reproduction number (Rt) to be 2.70, 95% HPD (1.76, 4.08) in Ontario and 2.10, 95% HPD (1.41, 3.17) in Maryland. The Rt trends in Ontario closely matched those derived via EpiEstim using reported case numbers. Our study also provides new insights into the median infection duration of EV-D68, estimated at 7.94 days, 95% HPD (4.55, 12.8) in Ontario and 10.8 days, 95% HPD (5.85, 18.6) in Maryland, addressing the gap in the existing literature surrounding EV-D68’s infection period. We observed that the estimated Time since the Most Recent Common Ancestor (TMRCA) and the epidemic’s origin coincided with the easing of COVID-19 related social contact restrictions in both areas. This suggests that the relaxation of non-pharmaceutical interventions, initially implemented to control COVID-19, may have inadvertently facilitated the spread of EV-D68. These findings underscore the effectiveness of phylodynamic methods in public health, demonstrating their broad application from local to global scales and underscoring the critical role of pathogen genomic data in enhancing public health surveillance and outbreak characterization.

Abstract Background For accessing dental care in Canada, approximately 62% of the population has employment-based insurance, 6% have some publicly funded coverage, and 32% have to pay out-of pocket. Those with no insurance or public coverage find dental care more unaffordable compared to those with private insurance. To support the development of more comprehensive publicly funded dental care programs, it is important to understand the socio-demographic attributes of all those, who find dental care unaffordable. Methods This study is a secondary analysis of the data collected from Ontarians during the latest available cycle of the Canadian Community Health Survey (2017-18), a cross-sectional survey that collects information on health status, health care utilization, and health determinants for the Canadian population. First, bivariate analysis was conducted to determine the characteristics of Ontarians who lack dental insurance. Afterwards, we employed machine learning (ML) to analyze data and identify risk indicators for not having private dental insurance. Specifically, we trained several supervised ML models and utilized Shapley additive explanations (SHAP) to determine the relative feature importance for not having private dental insurance from the best ML model [the gradient boosting (GBM)]. Results Approximately one-third of Ontarians do not have private insurance coverage for dental care. Individuals with an income below $20,000, those unemployed or working part-time, seniors aged above 70, and those unable to afford to have their own housing are more at risk of not having private dental insurance, leading to financial barriers in accessing dental care. Conclusion In the future, government-funded programs can incorporate these identified risk indicators when determining eligible populations for publicly funded dental programs. Understanding these attributes is critical for developing targeted and effective interventions, ensuring equitable access to dental care for Canadians.

Abstract Proteasome subunit hRpn13 is partially proteolyzed in certain cancer cell types to generate hRpn13Pru by degradation of its UCHL5/Uch37-binding DEUBAD domain and retention of an intact proteasome- and ubiquitin-binding Pru domain. By using structure-guided virtual screening, we identify an hRpn13 binder (XL44) and solve its structure ligated to hRpn13 Pru by integrated X-ray crystallography and NMR to reveal its targeting mechanism. Surprisingly, hRpn13Pru is depleted in myeloma cells following treatment with XL44. TMT-MS experiments reveal a select group of off-targets, including PCNA clamp-associated factor PCLAF and ribonucleoside-diphosphate reductase subunit M2 (RRM2), that are similarly depleted by XL44 treatment. XL44 induces hRpn13-dependent apoptosis and also restricts cell viability by a PCLAF-dependent mechanism. A KEN box, but not ubiquitination, is required for XL44-induced depletion of PCLAF. Here, we show that XL44 induces ubiquitin-dependent loss of hRpn13Pru and ubiquitin-independent loss of select KEN box containing proteins.

Blast-related traumatic brain injury (bTBI) is a major cause of neurological disorders in the U.S. military that can adversely impact some civilian populations as well and can lead to lifelong deficits and diminished quality of life. Among these types of injuries, the long-term sequelae are poorly understood because of variability in intensity and number of the blast exposure, as well as the range of subsequent symptoms that can overlap with those resulting from other traumatic events (e.g., post-traumatic stress disorder). Despite the valuable insights that rodent models have provided, there is a growing interest in using injury models using species with neuroanatomical features that more closely resemble the human brain. With this purpose, we established a gyrencephalic model of blast injury in ferrets, which underwent blast exposure applying conditions that closely mimic those associated with primary blast injuries to warfighters. In this study, we evaluated brain biochemical, microstructural, and behavioral profiles after blast exposure using in vivo longitudinal magnetic resonance imaging, histology, and behavioral assessments. In ferrets subjected to blast, the following alterations were found: 1) heightened impulsivity in decision making associated with pre-frontal cortex/amygdalar axis dysfunction; 2) transiently increased glutamate levels that are consistent with earlier findings during subacute stages post-TBI and may be involved in concomitant behavioral deficits; 3) abnormally high brain N-acetylaspartate levels that potentially reveal disrupted lipid synthesis and/or energy metabolism; and 4) dysfunction of pre-frontal cortex/auditory cortex signaling cascades that may reflect similar perturbations underlying secondary psychiatric disorders observed in warfighters after blast exposure.

Abstract Here, an in vitro characterization of a family of prazole derivatives that covalently bind to the C73 site on Tsg101 and assay their ability to inhibit viral particle production is presented. Structurally, increased steric bulk on the 4‐pyridyl of the prazole expands the prazole site on the UEV domain toward the β‐hairpin in the Ub‐binding site and is coupled to increased inhibition of virus‐like particle production in HIV‐1. Increased bulk also increased toxicity, which is alleviated by increasing flexibility. Further, the formation of a novel secondary Tsg101 adduct for several of the tested compounds and the commercial drug lansoprazole. The secondary adduct involved the loss of the 4‐pyridyl substituent to form an irreversible species, with implications for increasing the half‐life of the active species or its specificity toward Tsg101 UEV. It is also determined that sulfide derivatives display effective viral inhibition, presumably through cellular sulfoxidation, allowing for delayed conversion within the cellular environment, and identify SARS‐COV‐2 as a target of prazole inhibition. These results open multiple avenues for the design of prazole derivatives for antiviral applications.

Technological advancements in phylodynamic modeling coupled with the accessibility of real-time pathogen genetic data are increasingly important for understanding the infectious disease transmission dynamics. In this study, we compare the transmission potentials of North American influenza A(H1N1)pdm09 derived from sequence data to that derived from surveillance data. The impact of the choice of tree-priors, informative epidemiological priors, and evolutionary parameters on the transmission potential estimation is evaluated. North American Influenza A(H1N1)pdm09 hemagglutinin (HA) gene sequences are analyzed using the coalescent and birth-death tree prior models to estimate the basic reproduction number (R0). Epidemiological priors gathered from published literature are used to simulate the birth-death skyline models. Path-sampling marginal likelihood estimation is conducted to assess model fit. A bibliographic search to gather surveillance-based R0 values were consistently lower (mean ≤ 1.2) when estimated by coalescent models than by the birth-death models with informative priors on the duration of infectiousness (mean ≥ 1.3 to ≤2.88 days). The user-defined informative priors for use in the birth-death model shift the directionality of epidemiological and evolutionary parameters compared to non-informative estimates. While there was no certain impact of clock rate and tree height on the R0 estimation, an opposite relationship was observed between coalescent and birth-death tree priors. There was no significant difference (p = 0.46) between the birth-death model and surveillance R0 estimates. This study concludes that tree-prior methodological differences may have a substantial impact on the transmission potential estimation as well as the evolutionary parameters. The study also reports a consensus between the sequence-based R0 estimation and surveillance-based R0 estimates. Altogether, these outcomes shed light on the potential role of phylodynamic modeling to augment existing surveillance and epidemiological activities to better assess and respond to emerging infectious diseases.

Objective: To assess public awareness regarding endodontic treatment and assess patients' knowledge regarding endodontic treatment. Materials and Methods: A questionnaire was prepared and given to 300 patients who had visited the Department of Conservative Dentistry and Endodontics between November 2021 and October 2022, after obtaining ethical clearance and consent from all the participants. The questionnaire included sociodemographic details and about their knowledge about endodontic treatment. The collected data were tabulated and analyzed. Statistical Analysis: Data analysis was performed using methods of descriptive statistics like frequency and percentages. Results: We found that most of the respondents had an average level of knowledge regarding endodontic treatment. Conclusion: We observed an improvement in knowledge and perception regarding endodontic treatment.

Rare variant association data from human genetics combined with in vitro and in vivo functional validation highlight ANGPTL7 as a promising therapeutic target for intraocular pressure reduction, and protection from glaucoma.

A global monkeypox outbreak began in May 2022. Limited data exist on specimen type performance in associated molecular diagnostics. Consequently, a diverse range of specimen sources were collected in the initial weeks of the outbreak in Ontario, Canada. Our clinical evaluation identified skin lesions as the optimal diagnostic specimen source.

Abstract Risk stratification for hospitalized adults with COVID-19 is essential to inform decisions about individual patients and allocation of resources. So far, risk models for severe COVID outcomes have included age but have not been optimized to best serve the needs of either older or younger adults. Models also need to be updated to reflect improvements in COVID-19 treatments. This retrospective study analyzed data from 6906 hospitalized adults with COVID-19 from a community health system across five states in the western United States. Risk models were developed to predict mechanical ventilation illness or death across one to 56 days of hospitalization, using clinical data available within the first hour after either admission with COVID-19 or a first positive SARS-CoV-2 test. For the seven-day interval, models for age ≥ 18 and

Real-time device-level multi-domain emulation can provide an accurate insight into behavioral transients of the hydrogen fuel-cell hybrid electric bus (HEB). However, the conventional electromagnetic transient (EMT) simulation suffers from the computation burden caused by the complex multi-domain subsystems. This paper develops a hybrid recurrent neural network (RNN) and EMT method for device-level multi-domain emulation for fuel-cell and battery HEB. Two recurrent neural networks (RNN) are designed and trained to create device-level models of permanent magnet synchronous motor (PMSM) and the modular multilevel converter (MMC), respectively. The IGBTs’ behavioral transients and thermal performance are integrated into the RNN-based MMC model. Moreover, the EMT models represent the energy behaviour of onboard fuel-cell stacks and battery stacks. The proposed multi-domain hybrid models are implemented on the Xilinx Versal™ adaptive compute acceleration platform (ACAP), where multiple AI engines and the programmable logic deal with the RNN and EMT models, respectively. The real-time hardware emulation is carried out at the time-step of 0.1 $\mu \text{s}$ for device-level transients. The results show that the hybrid model has 96.3% accuracy; furthermore, it significantly reduces the HEB emulation time compared to conventional EMT methods.

In this paper, a modulated dual-voltage-vector model-free predictive current control with online duty cycle calculation is proposed and applied to drive a synchronous reluctance motor. The dual-voltage-vector modulation scheme reduces the current ripples and errors in the single-voltage-vector method. The proposed method reduces the predictive current controller’s calculation time by first setting the initial value of the duty cycle to a constant. Then, an optimal switching mode can be selected by minimizing a cost function. Next, the required duty cycle can be calculated directly by the proposed method without any differential calculations instead of its initial value. The proposed method can effectively track the stator current, reducing the maximum average current error by 34.8% compared to the conventional single-voltage-vector scheme. Finally, the correctness and feasibility of the modulated model-free predictive current controller with the online duty cycle calculation proposed in this article are verified by the experimental results using Texas Instruments microcontroller TMS320F28379D.

Abstract Herein, we report the theoretical investigation on the photonic nanojets (PNJs) of single dielectric microspheres illuminated by focused broadband radiation (polychromatic light) from a Halogen lamp, supercontinuum source, light-emitting diode, and Hg arc lamp. The role of incident beam waist, refractive index of the surrounding medium, and radius of the microsphere on the characteristic parameters such as the electric field intensity enhancement, effective width, and length of the PNJ is studied. Interestingly, the characteristic parameters of the PNJs of solid microspheres obtained for the above-mentioned broadband radiation sources are found close to those observed for the focused monochromatic radiation of wavelengths which are near to the central wavelengths of the sources. Moreover, the characteristic parameters of PNJs of the core–shell microspheres of different thicknesses (t) illuminated by polychromatic radiation from most commonly used sources such as Halogen and Hg arc lamps are studied. For each t value, a suitable wavelength of monochromatic radiation has been found to generate the PNJ with characteristic parameters which are close to those obtained in the case of polychromatic radiation. We believe that the analytical theory and the theoretical simulations reported here would be useful for researchers who work in the fields such as PNJ assisted photoacoustic spectroscopy, white light nanoscopy, low-coherence phase-shifting interference microscopy, and Mirau interferometry.

Rpn13 is a substrate receptor of the 26S proteasome and an anti-cancer drug target. Here, the authors identify and characterize XL5, a lead compound that binds to the N-terminal Pru domain of human Rpn13 (hRpn13), solve the NMR structure of XL5-ligated hRpn13 Pru and develop XL5-PROTACs that preferentially target an identified hRpn13 Pru fragment present in multiple myeloma cells.

The iRGD tumor-penetrating peptide can achieve tumor specific drug delivery but whether and how it can penetrate into desmoplastic tumors is unknown. Here, the authors show that β5 integrin expression on tumor cells, mediated by CAFs-derived TGF-β, is required for iRGD penetration into the desmoplastic PDAC microenvironment and that iRGD-based combination therapy is effective in PDAC mouse models.

Hydatid disease, is an infection caused by the larval stage (metacestode) of the cestode Echinococcus granulosus. In humans it may result in a wide spectrum of clinical manifestations ranging from asymptomatic infection to severe, even fatal disease. Echinococcal cysts are found in the liver in approximately 70 % of cases, and the lungs in approximately 25 % of cases. Cardiac hydatid disease is seen in 0.5–5% of total hydatid disease patients, the present case highlights the unusual presentation of Hydatid cyst in a different clinical scenario.

Abstract Prolactin (PRL) hormone functions as a pleiotropic cytokine with a protective role in the retina. We recently identified by transcriptome profiling that PRL is one of the most highly upregulated mRNAs in the retinas of mutant rcd1 (PDE6B) and xlpra2 (RPGR) dogs at advanced stages of photoreceptor disease. In the present study, we have identified the expression of a short PRL isoform that lacks exon 1 in canine retinas and analyzed the time-course of expression and localization of this isoform in the retinas of these two models. Using laser capture microdissection to isolate RNA from each of the retinal cellular layers, we found by qPCR that this short PRL isoform is expressed in photoreceptors of degenerating retinas. We confirmed by in situ hybridization that its expression is localized to the outer nuclear layer and begins shortly after the onset of disease at the time of peak photoreceptor cell death in both models. PRL protein was also detected only in mutant dog retinas. Our results call for further investigations into the role of this novel PRL isoform in retinal degeneration.

Real-time device-level hardware-in-the-loop (HIL) emulation of a complete high-speed rail system is challenging due to its complex modeling and high computing demand. With higher energy-efficient and switching frequency semiconductor material being found and adopted in the power electronic electrified traction application, there is a need to develop such new material based real-time device-level power electronic models in the modern and future traction system to estimate and verify the device switching transients, energy efficiency, and power quality improvement capability. This paper proposes a real-time SiC IGBT model based on the Wiener-Hammerstein configuration. A complete Beijing-Shanghai AC traction application is utilized as the study case, implemented on the hybrid multiprocessor system-on-chip (MPSoC) and field-programmable gate array (FPGA) platform, to verify both the system-level and device-level performance of the proposed model with comparisons to commercial software PSCAD/EMTDC and SaberRD. The dedicated hardware implementation enabled model execution at $10~ns$ for device-level transients and 10 μs for system-level transients.

Background: Angiotensin-converting enzyme inhibitors (ACEi) and angiotensin-II receptor blockers (ARB), the most commonly prescribed antihypertensive medications, counter renin-angiotensin-aldosterone system (RAAS) activation via induction of angiotensin-converting enzyme 2 (ACE2) expression. Considering that ACE2 is the functional receptor for SARS-CoV-2 entry into host cells, the association of ACEi and ARB with COVID-19 outcomes needs thorough evaluation. Methods: We conducted retrospective analyses using both unmatched and propensity score (PS)-matched cohorts on electronic health records (EHRs) to assess the impact of RAAS inhibitors on the risk of receiving invasive mechanical ventilation (IMV) and 30-day mortality among hospitalized COVID-19 patients. Additionally, we investigated the immune cell gene expression profiles of hospitalized COVID-19 patients with prior use of antihypertensive treatments from an observational prospective cohort. Results: The retrospective analysis revealed that there was no increased risk associated with either ACEi or ARB use. In fact, the use of ACEi showed decreased risk for mortality. Survival analyses using PS-matched cohorts suggested no significant relationship between RAAS inhibitors with a hospital stay and in-hospital mortality compared to non-RAAS medications and patients not on antihypertensive medications. From the analysis of gene expression profiles, we observed a noticeable up-regulation in the expression of 1L1R2 (an anti-inflammatory receptor) and RETN (an immunosuppressive marker) genes in monocytes among prior users of ACE inhibitors. Conclusion: Overall, the findings do not support the discontinuation of ACEi or ARB treatment and suggest that ACEi may moderate the COVID-19 hyperinflammatory response.

Genetic studies must enroll large numbers of participants to obtain adequate statistical power. Data are needed on how researchers can best use limited financial and practical resources to achieve these targets, especially in under-represented populations. This paper provides a retrospective analysis of the recruitment strategies for a large glaucoma genetics study in African Americans. The Primary Open-Angle African American Glaucoma Genetics study enrolled 10,192 African American subjects from the Philadelphia region. Major recruitment approaches included clinic enrollment from University of Pennsylvania (UPenn) sites, clinic enrollment from external sites, sampling of Penn Medicine Biobank (PMBB), and community outreach. We calculated the enrollment yield, cost per subject, and seasonal trends of these approaches. The majority (65%) of subject were enrolled from UPenn sites with an average cost of $133/subject. Over time, monthly case enrollment declined as the pool of eligible subjects was depleted. Expanding to external sites boosted case numbers ($129/subject) and the biobank provided additional controls at low cost ($5/subject), in large part due to the generosity of PMBB providing samples free of cost. Community outreach was costly with low return on enrollment ($978/subject for 220 subjects). Summer months (Jun-Aug) produced the highest recruitment yields (p

Introduction. Previous COVID-19 prognostic models have been developed in hospital settings and are not applicable to COVID-19 cases in the general population. There is an urgent need for prognostic scores aimed to identify patients at high risk of complications at the time of COVID-19 diagnosis. Methods. The RDT COVID-19 Observational Study (RCOS) collected clinical data from patients with COVID-19 admitted regardless of the severity of their symptoms in a general hospital in India. We aimed to develop and validate a simple bedside prognostic score to predict the risk of hypoxaemia or death. Results. 4035 patients were included in the development cohort and 2046 in the validation cohort. The primary outcome occurred in 961 (23.8%) and 548 (26.8%) patients in the development and validation cohorts, respectively. The final model included 12 variables: age, systolic blood pressure, heart rate, respiratory rate, aspartate transaminase, lactate dehydrogenase, urea, C-reactive protein, sodium, lymphocyte count, neutrophil count, and neutrophil/lymphocyte ratio. In the validation cohort, the area under the receiver operating characteristic curve (AUROCC) was 0.907 (95% CI, 0.892–0.922), and the Brier Score was 0.098. The decision curve analysis showed good clinical utility in hypothetical scenarios where the admission of patients was decided according to the prognostic index. When the prognostic index was used to predict mortality in the validation cohort, the AUROCC was 0.947 (95% CI, 0.925–0.97) and the Brier score was 0.0188. Conclusions. The RCOS prognostic index could help improve the decision making in the current COVID-19 pandemic, especially in resource-limited settings with poor healthcare infrastructure such as India. However, implementation in other settings is needed to cross-validate and verify our findings.

Abstract Prostate cancer (PC) is the sixth most common cancer type in the world, which causes approximately 10% of total cancer fatalities. The detection of protein biomarkers in body fluids is the key topic for the diagnosis and prognosis of PC. Highly sensitive screening of PC is the most effective approach for reducing mortality. Thus, there are a growing number of literature that recognizes the importance of new technologies for early diagnosis of PC. Graphene is playing an important role in the biosensor field with remarkable physical, optical, electrochemical and magnetic properties. Many recent studies demonstrated the potential of graphene materials for sensitive detection of protein biomarkers. In this review, the graphene-based biosensors toward PC analysis are mainly discussed in two groups: Firstly, novel biosensor interfaces were constructed through the modification of graphene materials onto sensor surfaces. Secondly, ingenious signal amplification strategies were developed using graphene materials as catalysts or carriers. Graphene-based biosensors have exhibited remarkable performance with high sensitivities, wide detection ranges, and long-term stabilities.

Abstract The enzyme succinate dehydrogenase (SDH) functions in the citric acid cycle and loss of function predisposes to the development of phaeochromocytoma/paraganglioma (PPGL), wild type gastrointestinal stromal tumour (wtGIST) and renal cell carcinoma. SDH-deficient tumours are most commonly associated with a germline SDH subunit gene (SDHA/B/C/D) mutation but can also be associated with epigenetic silencing of the SDHC gene. However, clinical diagnostic testing for an SDHC epimutation is not widely available. The objective of this study was to investigate the indications for and the optimum diagnostic pathways for the detection of SDHC epimutations in clinical practice. SDHC promoter methylation analysis of 32 paraffin embedded tumours (including 15 GIST and 17 PPGL) was performed using a pyrosequencing technique and correlated with SDHC gene expression. SDHC promoter methylation was identified in 6 (18.7%) tumours. All 6 SDHC epimutation cases presented with SDH deficient wtGIST and 3/6 cases had multiple primary tumours. No case of constitutional SDHC promoter hypermethylation was detected. Whole genome sequencing of germline DNA from three wtGIST cases with an SDHC epimutation, did not reveal any causative sequence anomalies. Herein, we recommend a diagnostic workflow for the detection of an SDHC epimutation in a service setting.

This paper focuses on identifying crash risk factors associated with injury severity of teen drivers. Crash data obtained from the Highway Safety and Information System (HSIS) for the entire state of North Carolina, for years 2011 to 2013, was used for analysis and modeling. Among all the crashes during the study period, a total of 62,990 crashes involving teen drivers (15 to 19 years) were analyzed. A partial proportionality odds model was developed to identify factors contributing to injury severity of teen drivers. The results obtained indicate that teen drivers driving sports utility vehicles and pickup trucks are more likely to be severely injured when compared to teen drivers driving passenger cars. Teen drivers are more likely to be severely injured on weekdays, particularly during peak hours. The chances of teen drivers getting involved in severe injury crashes on Tuesdays and Fridays is higher when compared to Sundays. Age, gender, road configuration, terrain, adverse weather condition, and access control are observed to have a significant effect on teen driver's injury severity. Keywords: Teen drivers, Crash, Injury severity, Contributing factor, Partial proportionality odds model

Abstract Background Age-related macular degeneration (AMD) is a degenerative disorder of the central retina and the foremost cause of blindness. The retinal pigment epithelium (RPE) is a primary site of disease pathogenesis. The genetic basis of AMD is relatively well understood; however, this knowledge is yet to yield a treatment for the most prevalent non-neovascular disease forms. Therefore, tissue-specific epigenetic mechanisms of gene regulation are of considerable interest in AMD. We aimed to identify differentially methylated genes associated with AMD in the RPE and differentiate local DNA methylation aberrations from global DNA methylation changes, as local DNA methylation changes may be more amenable to therapeutic manipulation. Methods Epigenome-wide association study and targeted gene expression profiling were carried out in RPE cells from eyes of human donors. We performed genome-wide DNA methylation profiling (Illumina 450k BeadChip array) on RPE cells from 44 human donor eyes (25 AMD and 19 normal controls). We validated the findings using bisulfite pyrosequencing in 55 RPE samples (30 AMD and 25 normal controls) including technical (n = 38) and independent replicate samples (n = 17). Long interspersed nucleotide element 1 (LINE-1) analysis was then applied to assess global DNA methylation changes in the RPE. RT-qPCR on independent donor RPE samples was performed to assess gene expression changes. Results Genome-wide DNA methylation profiling identified differential methylation of multiple loci including the SKI proto-oncogene (SKI) (p = 1.18 × 10−9), general transcription factor IIH subunit H4 (GTF2H4) (p = 7.03 × 10−7), and Tenascin X (TNXB) (p = 6.30 × 10−6) genes in AMD. Bisulfite pyrosequencing validated the differentially methylated locus cg18934822 in SKI, and cg22508626 within GTF2H4, and excluded global DNA methylation changes in the RPE in AMD. We further demonstrated the differential expression of SKI, GTF2H4, and TNXB in the RPE of independent AMD donors. Conclusions We report the largest genome-wide methylation analysis of RPE in AMD along with associated gene expression changes to date, for the first-time reaching genome-wide significance, and identified novel targets for functional and future therapeutic intervention studies. The novel differentially methylated genes SKI and GTF2H4 have not been previously associated with AMD, and regulate disease pathways implicated in AMD, including TGF beta signaling (SKI) and transcription-dependent DNA repair mechanisms (GTF2H4).

Background Coronavirus disease 2019 (COVID-19) is associated with endothelial activation and coagulopathy, which may be related to pre-existing or infection-induced pro-thrombotic autoantibodies such as those targeting angiotensin II type I receptor (AT1R-Ab). Methods We compared prevalence and levels of AT1R-Ab in COVID-19 cases with mild or severe disease to age and sex matched negative controls utilizing multivariate logistic and quantile regression adjusted for comorbidities including hypertension, diabetes, and heart disease. Results There were trends toward increased prevalence (50% vs. 33%, p = 0.1) and level of AT1R-Ab (median 9.8 vs. 6.1 U/mL, p = 0.06) in all cases versus controls. When considered by COVID-19 disease severity, there was a trend toward increased prevalence of AT1R-Ab (55% vs. 31%, p = 0.07), as well as significantly higher AT1R-Ab levels (median 10.7 vs. 5.9 U/mL, p = 0.03) amongst individuals with mild COVID-19 versus matched controls. In contrast, the prevalence (42% vs. 37%, p = 0.9) and level (both medians 6.7 U/mL, p = 0.9) of AT1R-Ab amongst those with severe COVID-19 did not differ from matched controls. Conclusions These findings support an association between COVID-19 and AT1R-Ab, emphasizing that vascular pathology may be present in individuals with mild COVID-19 as well as those with severe disease.

Single-vehicle run-off-road (ROR) crashes account for a significant number of all the single-vehicle crashes and all the fatalities in the United States. The variable categories influencing single-vehicle ROR crashes and their severity could be different from other single-vehicle crashes. This paper, therefore, focuses on investigating single-vehicle ROR crashes to identify and compare variable categories influencing their occurrence and severity. A logistic regression model was developed to evaluate and identify variable categories associated with single-vehicle ROR crashes compared to other single-vehicle crashes. The results indicate that variables such as road characteristics (number of lanes, road surface condition, road terrain, functional classification, and geometric condition), weather condition, road lighting condition, vehicle type, and, driver characteristics (driver physical condition, age, and gender) are observed to have a statistically significant influence on the occurrence of single-vehicle ROR crashes. A partial proportionality odds model was developed to evaluate and identify variable categories associated with injury severity of single-vehicle ROR crashes. The results indicate that variables and variable categories such as weather condition (cloudy and rain), speed limit ( 88.5 kmph [55 mph]), vehicle type (primarily buses), drivers’ age (

Fog and rain influence the ability of the drivers to see other vehicles, pedestrians, bicyclists, and fixed objects, and, hence, increase the likelihood of getting involved in a crash on a road. Resource constraints limit installing weather monitoring sensors at regular intervals (for example, every 1, 2, 3, or 4 km) on a road. Therefore, the objective of this paper is to develop a feed-forward back-propagation neural network (BPNN) model to predict fog / low visibility condition, and, apply at a road link-level. Five years of meteorological data were collected from 238 weather monitoring stations in and around North Carolina to develop and validate the BPNN visibility model. The predicted visibility using meteorological data was compared with visibility from the weather monitoring sensor installed at five geographically distributed locations in North Carolina. The BPNN visibility model has better visibility prediction capability during a normal foggy condition when compared to visibility during thick/dense fog condition.

The effect of a planned special event on travel time performance measures varies spatially and temporally based on the type of the planned special event. Evaluating and assessing the effect helps practitioners proactively plan and improve traffic operations within its vicinity during the future planned special event. The objective of this paper, therefore, is to adopt a systemic evaluation method and examine the effect of a planned special event on travel time performance measures, over time and space, from the location of the planned special event. Raw travel time data obtained from a private data source was used to assess the spatial and temporal effects of two planned special events on travel time performance measures. Data were captured for each major road link within 3-miles of each planned special event location. Statistical analyses such as the t-test and Cohen's D were conducted to assess the difference in travel time performance measures during the day of the planned special event compared to the normal day. A statistically significant increase in travel times and travel time variations was observed on the day of the planned special event when compared to the normal day. However, the effect of the increase in travel times was observed to be small, whereas, the effect of the increase in the travel time variation was observed to be higher. Buffer time index (BTI) and planning time index (PTI) were observed to be significantly higher during the NASCAR race and for longer periods compared to the NFL game.

The 2014-2015 highly pathogenic avian influenza (HPAI) H5NX outbreak represents the largest and most expensive HPAI outbreak in the United States to date. Despite extensive traditional and molecular epidemiological studies, factors associated with the spread of HPAI among midwestern poultry premises remain unclear. To better understand the dynamics of this outbreak, 182 full genome HPAI H5N2 sequences isolated from commercial layer chicken and turkey production premises were analyzed using evolutionary models able to accommodate epidemiological and geographic information. Epidemiological compartmental models embedded in a phylogenetic framework provided evidence that poultry type acted as a barrier to the transmission of virus among midwestern poultry farms. Furthermore, after initial introduction, the propagation of HPAI cases was self-sustainable within the commercial poultry industries. Discrete trait diffusion models indicated that within state viral transitions occurred more frequently than inter-state transitions. Distance and sample size were very strongly supported as associated with viral transition between county groups (Bayes Factor > 30.0). Together these findings indicate that the different types of midwestern poultry industries were not a single homogenous population, but rather, the outbreak was shaped by poultry industries and geographic factors.

The objective of this research is to identify factors associated with crashes due to overcorrection or oversteering of vehicles. Crash data was collected from 2011 to 2013 for the State of North Carolina in the United States. Logistic regression modeling was used to analyze crash data because of the dichotomous nature of the dependent variable (overcorrection or oversteering). The crash involvement due to overcorrection or oversteering of a vehicle decreased as the age of the driver increased. Drivers are 2.22 times more likely to overcorrect or oversteer when ill, 3.44 times more likely to overcorrect or oversteer when under fatigue, and 1.61 times more likely to overcorrect or oversteer when fallen asleep compared to normal physical conditions. Overall, driver characteristics and speed limit tend to play a major role in overcorrection or oversteering of vehicles. Programs to reduce impaired driving might help in the reduction of overcorrection or oversteering related crash fatalities or injuries. Additionally, training and driver education programs focusing on identified factors associated with crashes due to overcorrection or oversteering of vehicles will benefit drivers on how to respond during emergency or panic situations. Keywords: Overcorrection, Oversteering, Vehicle, Logistic regression, Crash

Objective: To evaluate the quantitative expression of Ki-67 in oral leukoplakia, oral submucous fibrosis (OSF) and to compare their expression with normal oral epithelium. Materials and Methods: Thirty archival paraffin-embedded blocks were retrieved from the Department of Oral and Maxillofacial Pathology. It includes three groups with 10 specimens each of histopathologically confirmed oral leukoplakia, OSF, and controls, which were taken as standard for comparison. Ki-67 immunostaining was detected and numbers of positive cells out of 1000 were counted in 10 random high-power fields in each case. Statistical analysis was done using one-way analysis of variance test. Results: Mean and standard deviation of oral leukoplakia was found to be higher (398 ± 82) followed by OSF (343 ± 83) and controls (290 ± 72). Pairwise comparison of three groups with respect to expression of total number and percentage of cells was statistically significant between oral leukoplakia and controls (P < 0.05). Conclusion: The results of the present study emphasize that malignant transformation rate was higher for oral leukoplakia compared with OSF and controls. Hence, it serves as a prognostic tool for early detection of malignancy.

Background: Oral sub mucous fibrosis (OSMF) is chronic insidious diseases of the oral cavity, pharynx and larynx with unknown etiology. It is characterized by mucosal rigidity. The proposed etiological factor for causing OSMF areca nut. Areca nut and arecoline cause fibroblast proliferation and collagen production. Aim and Objectives: Correlate the clinical and histopathological grading using Masson's trichrome and Van Gieson's stain in OSMF. Materials and Methods: Thirty patients with clinically proven cases of OSMF and grouped according to their mouth opening. Incisional biopsy was done. The sections were stained with hematoxylin and eosin for histopathological conformation. Selected cases were sectioned and stained with Masson's trichrome and Van Gieson special stain and graded histopathologically. Data was analyzed using SPSS 20.0 version. Pearson's correlation coefficient and Chi-square test were done to find out the association between variables. Student's t-test was used to find clinical and histopathological correlation. Results: The mean age of OSMF patients was 20–40 years and when compared with gender males were affected more than females. The stains between Masson's trichrome with Van Gieson's stain showed significant correlation, but when compared between clinical and histopathological grading there was no statistically significant. Conclusion: Clinical staging and histopathological grading in OSMF there is no correlation when using special stains such as Masson's trichrome and Van Gieson's stain.

We intend to identify marker genes with differential gene expression (DEG) and RGC subtypes in cultures of human-induced pluripotent stem cell (iPSC)-derived retinal ganglion cells. Single-cell sequencing was performed on mature and functional iPSC-RGCs at day 40 using Chromium Single Cell 3’ V3 protocols (10X Genomics). Sequencing libraries were run on Illumina Novaseq to generate 150 PE reads. Demultiplexed FASTQ files were mapped to the hg38 reference genome using the STAR package, and cluster analyses were performed using a cell ranger and BBrowser2 software. QC analysis was performed by removing the reads corresponding to ribosomal and mitochondrial genes, as well as cells that had less than 1X mean absolute deviation (MAD), resulting in 4705 cells that were used for further analyses. Cells were separated into clusters based on the gene expression normalization via PCA and TSNE analyses using the Seurat tool and/or Louvain clustering when using BBrowser2 software. DEG analysis identified subsets of RGCs with markers like MAP2, RBPMS, TUJ1, BRN3A, SOX4, TUBB3, SNCG, PAX6 and NRN1 in iPSC-RGCs. Differential expression analysis between separate clusters identified significant DEG transcripts associated with cell cycle, neuron regulatory networks, protein kinases, calcium signaling, growth factor hormones, and homeobox transcription factors. Further cluster refinement identified RGC diversity and subtype specification within iPSC-RGCs. DEGs can be used as biomarkers for RGC subtype classification, which will allow screening model systems that represent a spectrum of diseases with RGC pathology.

We investigated the association of the single nucleotide polymorphism (SNP) rs112369934 near the TRIM66 gene with qualitative and quantitative phenotypes of primary open-angle glaucoma (POAG) in African Americans (AA). AA subjects over 35 years old were recruited for the Primary Open-Angle African American Glaucoma Genetics (POAAGG) study in Philadelphia, PA. Glaucoma cases were evaluated for phenotypes associated with POAG pathogenesis, and the associations between rs112369934 and phenotypes were investigated by logistic regression analysis and in gender-stratified case cohorts: The SNP rs112369934 was found to have a suggestive association with retinal nerve fiber layer (RNFL) thickness and cup-to-disc ratio (CDR) in 1087 male AA POAG cases, individuals with the TC genotype having thinner RNFL (95% CI 0.85 to 6.61, p = 0.01) and larger CDR (95% CI −0.07 to −0.01, p = 0.02) than those with wildtype TT. No other significant associations were found. In conclusion SNP rs112369934 may play a role in POAG pathogenesis in male AA individuals. However, this SNP has been implicated in higher POAG risk in both male and female AA POAG cases.

Primary open-angle glaucoma (POAG) is the leading cause of irreversible blindness worldwide and has been associated with multiple genetic risk factors. The LMX1B gene is a genetic susceptibility factor for POAG, and several single-nucleotide polymorphisms (SNPs) were shown to be associated with POAG in our own prior Primary Open-Angle African American Glaucoma Genetics (POAAGG) study genome-wide association study (GWAS). This study evaluated the association of the LMX1B locus with baseline optic disc and clinical phenotypic characteristics of glaucoma patients from our African American cohort. Compared to the GG genotype in SNP rs187699205, the GC genotype in this SNP was found to be significantly associated with a smaller cup-to-disc ratio (CDR) and increased (better) visual field mean deviation (MD) in glaucoma cases. None of the glaucoma cases with the GC genotype had disc hemorrhages, disc notching, or beanpot disc appearance. In conclusion, glaucoma phenotypes differed significantly by LMX1B variant in African American patients with POAG, and a SNP variant was associated with certain disease features considered lower risk.

Introduction: Hookworm infection is a leading cause of iron deficiency anemia and malnutrition in resource-poor settings. Periodic mass deworming with anthelminthic drugs remains the cornerstone of hookworm control efforts worldwide. Reinfection following treatment occurs, reflecting the human host's inability to acquire immunity following exposure to an untreated reservoir of infection. This cluster randomized trial will evaluate the effectiveness of a modified, population-based, mass deworming strategy in reducing hookworm infection in an endemic southern Indian population. Methods: Forty five tribal villages were randomized into three groups: one received annual treatment; the second received two rounds of treatment at 1-month intervals; and the third received four rounds of treatment – two rounds 1 month apart at the beginning, followed by another two after 6 months. Stool samples collected through cross-sectional parasitological surveys pre- and post-intervention, and at 3-monthly intervals for a period of 1 year were tested for presence of hookworm ova. Long-term effectiveness of treatment will be assessed through another survey conducted 2 years after the last treatment cycle. Results: From a population of 11,857 individuals, 8681 (73.2%) were found to be eligible and consented to participate, out-migration being the primary reason for non-participation. Baseline stool samples were obtained from 2082 participants, with 18.5% having hookworm infection, although majority were low intensity infections (

Data from weather stations at airports, far away locations or predictions using macro-level data may not be accurate enough to disseminate visibility related information to motorists in advance. Therefore, the objective of this research is to investigate the influence of contributing factors and develop visibility prediction models, at road link-level, by considering data from weather stations located within 1.6 km of state routes, US routes and interstates in the state of North Carolina (NC). Four years of meteorological data, from January 2011 to December 2014, were collected within NC. Ordinary least squares (OLS) and weighted least squares (WLS) regression models were developed for different visibility and elevation ranges. The results indicate that elevation and cloud cover are negatively associated with low visibility. The chances of low visibility are higher between six to twelve hours after rainfall when compared to the first six hours after rainfall. A visibility sensor was installed at four different locations in NC to compare hourly visibility from the selected regression model, High-Resolution Rapid Refresh (HRRR) data, and the nearest weather station. The results indicate that the number of samples with zero error range was higher for the selected regression model compared with the HRRR and weather station observations.

High soil salinity is one of the most important environmental stressors, and is an increasing concern under the alarmingly global climate uncertainties. Salinity affects the growth and productivity of food crops, including rice, and ultimately threatens global food security and sustainability. Conventional breeding efforts to develop salt tolerant rice varieties utilizing its primary gene pool have resulted in limited success due to the mutligenic inheritance of salt tolerance traits. On the other hand, Porteresia coarctata, the only halophyte wild rice, offers tremendous opportunities to improve salt tolerance in rice. However, reports on the characterization of its rich genetic resources are scanty. The present study was undertaken to functionally identify salt responsive genes of P. coarctata in Arabidopsis model using a cDNA overexpression (COX) strategy. COX-mediated hunting of transgenic Arabidopsis expressing a cDNA library of P. coarctata under control of a maize ubiquitin promoter identified 12 candidates that were involved in salt tolerance for seed germination, and seedling and reproductive growth. Of these, four genes coding for a metallothionein, a ribosomal protein, a photosystem II 10 kDa protein, and a ferredoxin:thioredoxin reductase conferred enhanced salt tolerance phenotypes, such as better root and shoot growth, and tissue tolerance by maintaining higher relative water content, membrane stability and protecting cells from reactive oxygen species. These genes could be ideal candidates for improving salt tolerance in rice; however, further studies are needed to ascertain their direct functional relevance in salt tolerance mechanisms. Keywords: cDNA overexpression, Functional screening, halophyte, Porteresia coarctata salinity

Background Available outcomes data for anticoagulation therapy in adults with congenital heart disease (CHD) provide assessment of global risk of this therapy for CHD patients (a heterogeneous population), but the risk of complications for the different CHD diagnoses is unknown. The purpose of the study was to describe the indications for anticoagulation, and the incidence and risk factors for major bleeding complication in adults with tetralogy of Fallot. Methods and Results We queried Mayo Adult Congenital Heart Disease (MACHD) database for tetralogy of Fallot patients (aged ≥18 years) that received anticoagulation, 1990–2017. Of 130 patients (42±14 years, 75 men [58%]), warfarin and direct oral anticoagulants were used in 125 (96%) and 5 (4%), respectively because atrial arrhythmias (n=109), mechanical prosthetic valve (n=29), intracardiac thrombus (n=4), pulmonary embolism (n=6), stroke (n=3), and perioperative anticoagulation (n=44). The median hypertension, abnormal renal or liver function; stroke; bleeding history or predisposition; labile international normalized ratio; elderly (>65 years); drug or alcohol use score for the entire cohort was 1 (0–2) and 27 (21%) had hypertension, abnormal renal or liver function; stroke; bleeding history or predisposition; labile international normalized ratio; elderly (>65 years); drug or alcohol use score ≥2. There were 14 minor bleeding events (1.6% per year) and 11 major bleeding events (1.3% per year) in 8 patients during median follow‐up of 74 months (856 patient‐years). Mechanical prosthesis (hazard ratio 1.78, CI 1.29–3.77, P=0.021) and hypertension, abnormal renal or liver function; stroke; bleeding history or predisposition; labile international normalized ratio; elderly (>65 years); drug or alcohol use score ≥2 (hazard ratio 1.41, CI 1.03–3.88, P=0.046) were risk factors for major bleeding events. All‐cause mortality was higher in patients with major bleeding events (n=6, 75%) compared with patients without major bleeding events (n=25, 21%), P=0.001. Conclusions Considering the heterogeneity of the CHD population, data from the current study may be better suited for clinical decision‐making in tetralogy of Fallot patients.

The purpose of this study was to investigate the association between gender and primary open-angle glaucoma (POAG) among African Americans and to assess demographic, systemic, and behavioral factors that may contribute to differences between genders. The Primary Open-Angle African American Glaucoma Genetics (POAAGG) study had a case-control design and included African Americans 35 years and older, recruited from the greater Philadelphia, Pennsylvania. Diagnosis of POAG was based on evidence of both glaucomatous optic nerve damage and characteristic visual field loss. Demographic and behavioral information, history of systemic diseases and anthropometric measurements were obtained at study enrollment. Gender differences in risk of POAG were examined using multivariate logistic regression. A total of 2,290 POAG cases and 2,538 controls were included in the study. The percentage of men among cases was higher than among controls (38.6% vs 30.3%, P

Background Patients with tetralogy of Fallot (TOF) remain at risk for cardiovascular events despite successful repair. Some of the current risk stratification tools require advanced imaging and invasive studies, and hence are difficult to apply to routine patient care. A recent study showed that QRS fragmentation (QRS‐f) is predictive of mortality in patients with TOF. The current study aims to validate this result by assessing whether severity of QRS‐f could predict all‐cause mortality in a different TOF population. Methods and Results The authors reviewed the Mayo Adult Congenital Heart Disease database for patients with TOF who had ECG from 1990–2017. QRS‐f was defined as notches in QRS complex in ≥2 contiguous leads on ECG, not related to bundle branch block, and classified as none, mild (≤3 leads), moderate (4 leads), or severe (≥5 leads). Of 465 patients (age 37±14 years) in the study, QRS‐f was present in 161 (35%): mild (n=43, 9%), moderate (n=77, 17%), and severe (n=41, 9%). There were 55 deaths (12%) during 13.6±8.2 years of follow‐up. Severity of QRS‐f remained an independent predictor of all‐cause mortality after adjustment for other ECG parameters, patient demographics, and atrial and ventricular arrhythmia (hazard ratio, 1.74 per class; 95% confidence interval, 1.08–2.93 [P=0.041]). Conclusions The presence of severe QRS‐f may be used as complementary data to the usual clinical indices to determine whether interventions such as invasive electrophysiology study should be performed in patients with nonsustained ventricular tachycardia or to proceed with pulmonary valve replacement in patients with severe pulmonary regurgitation with ventricular volumes below the guideline‐directed threshold for intervention.

Sclerosing liposarcoma of epididymis is a rare extratesticular scrotal tumor with variable prognosis. Ultrasonography is the initial imaging modality of choice for the evaluation of scrotal mass and helps to differentiate testicular and extratesticular masses, thereby narrowing down the differential diagnosis. Magnetic resonance imaging with its excellent soft tissue resolution can help in the further characterization of the nature of the tumor. In this case report, we highlight the role of chemical shift imaging in making a confident preoperative diagnosis of liposarcoma thereby guiding optimal and timely management.

The genes in the 9p21 locus (CDKN2B-AS1 & CDKN2B) are widely associated with Primary open-angle glaucoma (POAG). However, the functional importance of this locus in POAG pathogenesis is still unexplored. This study investigated the role of CDKN2BAS1-CDKN2B axis in POAG. We observed significant association of CDKN2B-AS1 SNP rs4977756 with POAG and its endophenotypic traits (vertical cup-disc ratio (p = 0.033) and central corneal thickness (p = 0.008)) by screening African American POAG cases (n = 1567) and controls (n = 1600). A luciferase reporter assay in Human embryonic kidney 293T (HEK293T) cells revealed that the region surrounding rs4977756 likely serves as a transcriptional repressor. siRNA-mediated knockdown of CDKN2B-AS1 in HEK293T cells and trabecular meshwork (TM) cells resulted in significantly increased expression of CDKN2B, which was also observed in human POAG ocular tissues. Pathway focused qRT-PCR gene expression analysis showed increased cellular senescence, TGFβ signaling and ECM deposition in TM cells after CDKN2B-AS1 suppression. In conclusion, we report that CDKN2B-AS1 may act as a regulator, and it could function by modulating the expression of CDKN2B. In addition, increase in CDKN2B levels due to CDKN2B-AS1 suppression may result in the senescence of trabecular meshwork cells leading to POAG pathogenesis.

The world is combating a common and invisible enemy severe acute respiratory syndrome coronavirus 2 (SARS-CoV2), a highly transmissible virus responsible for serious respiratory illness coronavirus disease-2019 (COVID-19). As with all respiratory viruses, public health measures are focused on contact tracing, isolation, and treatment of affected individuals, who have respiratory symptoms. However, it is spreading efficiently, and it can be explained from its stealth transmission from presymptomatic and asymptomatic individuals. Droplet and contact precautions are followed universally. Healthcare workers are at higher risk of acquiring infection and they are additionally required to follow airborne and eye protection. Recent studies indicate viral particles can be isolated from many body fluids including feces, saliva, semen, and tears, suggesting transmission could be possibly occurring through some of these routes as well. We have done an evidence-based review of all potential modes of transmission and discussed preventive measures to stop the spread. There is an urgent need for educating the healthcare professionals, governments, and public regarding other potential modes of transmission. Strict preventive measures need to be used to stop the spread.