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Abstract Background The SARS-CoV-2 pandemic led to unprecedented testing demands, causing major testing delays globally. One strategy used for increasing testing capacity was pooled-testing, using a two-stage technique first introduced during WWII. However, such traditional pooled testing was used in practice only when positivity rates were below 2%. Methods Here we report the development, validation and clinical application of P-BEST - a single-stage pooled-testing strategy that was approved for clinical use in Israel. Results P-BEST is clinically validated using 3636 side-by-side tests and is able to correctly detect all positive samples and accurately estimate their Ct value. Following regulatory approval by the Israeli Ministry of Health, P-BEST was used in 2021 to clinically test 837,138 samples using 270,095 PCR tests - a 3.1fold reduction in the number of tests. This period includes the Alpha and Delta waves, when positivity rates exceeded 10%, rendering traditional pooling non-practical. We also describe a tablet-based solution that allows performing manual single-stage pooling in settings where liquid dispensing robots are not available. Conclusions Our data provides a proof-of-concept for large-scale clinical implementation of single-stage pooled-testing for continuous surveillance of multiple pathogens with reduced test costs, and as an important tool for increasing testing efficiency during pandemic outbreaks.

Abstract In 1974, Stephen Hawking predicted that quantum effects in the proximity of a black hole lead to the emission of particles and black hole evaporation. At the very heart of this process lies a logarithmic phase singularity which leads to the Bose-Einstein statistics of Hawking radiation. An identical singularity appears in the elementary quantum system of the inverted harmonic oscillator. In this Letter we report the observation of the onset of this logarithmic phase singularity emerging at a horizon in phase space and giving rise to a Fermi-Dirac distribution. For this purpose, we utilize surface gravity water waves and freely propagate an appropriately tailored energy wave function of the inverted harmonic oscillator to reveal the phase space horizon and the intrinsic singularities. Due to the presence of an amplitude singularity in this system, the analogous quantities display a Fermi-Dirac rather than a Bose-Einstein distribution.

The present study sought to examine the perceptions of teacher educators with respect to pedagogical innovation. The research included in-depth interviews with eighteen members of academic faculties at a teachers' education college. The findings revealed that teacher educators understand pedagogical innovation as having four central features: disrupting teaching regularities; listening to students' voices; promoting active learning based on personal responsibility; and, relevance in teaching-learning processes. Sustaining these features is the teacher educators' sense of commitment with respect to two facets: the first, to implement changes to teaching so as to adapt it to 21st century skills needed for the purpose of training and developing educators capable of working in a world of evolving realities; and, the second, to serve as a model of teaching for student teachers through direct and indirect modelling of these four features. A descriptive model relating the teacher educators' perceptions was constructed based on the research findings.

Antler is one of the primary animal raw materials exploited for technical purposes by the hunter-gatherer groups of the Eurasian Upper Palaeolithic (UP) all over the ecological range of deers, and beyond. It was exhaustively employed to produce one of the most critical tools for the survival of the UP societies: hunting weapons. However, antler implements can be made from diverse deer taxa, with different ecological requirements and ethological behaviours. Identifying the antler's origin at a taxonomic level is thus essential in improving our knowledge of humans' functional, practical and symbolic choices, as well as the human-animal interface during Prehistoric times. Nevertheless, palaeogenetics analyses have focused mainly on bone and teeth, with genetic studies of antler generally focused on modern deer conservation. Here we present the results of the first whole mitochondrial genome ancient DNA (aDNA) analysis by means of in-solution hybridisation capture of antlers from pre-Holocene archaeological contexts. We analysed a set of 50 Palaeolithic and Neolithic (c. 34-8ka) antler and osseous objects from South-Western Europe, Central Europe, South-Western Asia and the Caucasus. We successfully obtained aDNA, allowing us to identify the exploited taxa and demonstrate the archaeological relevance of those finds. Moreover, as most of the antlers were sampled using a minimally-invasive method, further analyses (morphometric, technical, genetic, radiometric and more) remain possible on these objects.

Abstract Circulating cell-free DNA (cfDNA) fragments are a biological analyte with extensive utility in diagnostic medicine. Understanding the source of cfDNA and mechanisms of release is crucial for designing and interpreting cfDNA-based liquid biopsy assays. Using cell type-specific methylation markers as well as genome-wide methylation analysis, we determine that megakaryocytes, the precursors of anuclear platelets, are major contributors to cfDNA (~26%), while erythroblasts contribute 1–4% of cfDNA in healthy individuals. Surprisingly, we discover that platelets contain genomic DNA fragments originating in megakaryocytes, contrary to the general understanding that platelets lack genomic DNA. Megakaryocyte-derived cfDNA is increased in pathologies involving increased platelet production (Essential Thrombocythemia, Idiopathic Thrombocytopenic Purpura) and decreased upon reduced platelet production due to chemotherapy-induced bone marrow suppression. Similarly, erythroblast cfDNA is reflective of erythrocyte production and is elevated in patients with thalassemia. Megakaryocyte- and erythroblast-specific DNA methylation patterns can thus serve as biomarkers for pathologies involving increased or decreased thrombopoiesis and erythropoiesis, which can aid in determining the etiology of aberrant levels of erythrocytes and platelets.

Diabetes Mellitus (DM) is a significant global concern. Many diabetic patients will experience complications due to angiopathy, neuropathy, and immune dysfunction, namely diabetic foot ulcers (DFU) and diabetic foot infections (DFI), which can result in lower limb amputation and potentially death. The prevalence of common superficial fungal infections, such as tinea pedis and onychomycosis, can directly increase a diabetic patient’s risk of developing both DFU and DFI. In this review article, we discuss the etiology of diabetic foot complications as well as considerations for both screening and management. We also discuss the role of the dermatologist within a multidisciplinary care team in prescribing and managing treatments for tinea pedis and onychomycosis infections within this patient population. We believe that reducing the burden of these fungal infections in the context of the diabetic foot will help reduce DFU and DFI complications and their associated morbidity and mortality.

Abstract Aim Understanding the association between pediatric feeding disorder (PFD) and age of presentation is limited. We aimed to investigate factors associated with PFD among different age groups. Methods Retrospective analysis of medical records of infants and toddlers diagnosed with PFD, according to the World Health Organization‐based definition. We compared children aged 1–12 months to those aged 13–72 months. Results Included were 253 children with PFD (median [interquartile range] age 16.4 [9.5–33] months at diagnosis, 56% boys). Significantly more children in the younger age group were girls (52.6% vs. 34.4%, respectively, p = .03) and preterm (25% vs. 14%, p = .03). They had more hospitalizations (34% vs. 23%, p = .03) and needed more prescription medications (36% vs. 17%, p

Objective To present a novel method that uses an epigenetic fingerprint to measure changes in plasma concentrations of cardiac-specific cell-free DNA (CS-cfDNA) as a marker of myocardial cell death. Methods This prospective, analytic, observational comparative study included patients with heart disease or multiple risk factors for heart disease undergoing major noncardiac, mostly vascular surgery, requiring an arterial-line, and at least 24 h hospitalization in the post anaesthesia care unit or critical care unit after surgery. Blood samples were collected at least four times per patient to measure troponin-T (via high-sensitivity troponin-T test) and CS-cfDNA pre- and postoperatively. Results A total of 117 patients were included (group 1, 77 patients [66%] with low preoperative and postoperative troponin-T; group 2, 18 patients [15%] with low preoperative but increased postoperative troponin-T; group 3, 16 patients [14%] with high troponin-T both preoperatively and postoperatively; and group 4, six patients [5%] with elevated preoperative troponin-T that decreased postoperatively). The increase in CS-cfDNA after surgery was statistically significant only in group 2, which correlated with an increase in troponin-T in the same group. Conclusions CS-cfDNA increased early postoperatively, particularly in patients with silent postoperative troponin elevation, and was correlated with an increase in troponin-T. These results may suggest that, in the subgroup of patients with postoperative elevated troponin, cardiomyocyte death indeed occurred.

Abstract Vaccination, especially with multiple doses, provides substantial population-level protection against COVID-19, but emerging variants of concern (VOC) and waning immunity represent significant risks at the individual level. Here we identify correlates of protection (COP) in a multicenter prospective study following 607 healthy individuals who received three doses of the Pfizer-BNT162b2 vaccine approximately six months prior to enrollment. We compared 242 individuals who received a fourth dose to 365 who did not. Within 90 days of enrollment, 239 individuals contracted COVID-19, 45% of the 3-dose group and 30% of the four-dose group. The fourth dose elicited a significant rise in antibody binding and neutralizing titers against multiple VOCs reducing the risk of symptomatic infection by 37% [95%CI, 15%-54%]. However, a group of individuals, characterized by low baseline titers of binding antibodies, remained susceptible to infection despite significantly increased neutralizing antibody titers upon boosting. A combination of reduced IgG levels to RBD mutants and reduced VOC-recognizing IgA antibodies represented the strongest COP in both the 3-dose group (HR = 6.34, p = 0.008) and four-dose group (HR = 8.14, p = 0.018). We validated our findings in an independent second cohort. In summary combination IgA and IgG baseline binding antibody levels may identify individuals most at risk from future infections.

BACKGROUND. Accurate detection of graft-versus-host disease (GVHD) is a major challenge in the management of patients undergoing hematopoietic stem cell transplantation (HCT). Here, we demonstrated the use of circulating cell-free DNA (cfDNA) for detection of tissue turnover and chronic GVHD (cGVHD) in specific organs. METHODS. We established a cocktail of tissue-specific DNA methylation markers and used it to determine the concentration of cfDNA molecules derived from the liver, skin, lungs, colon, and specific immune cells in 101 patients undergoing HCT. RESULTS. Patients with active cGVHD showed elevated concentrations of cfDNA, as well as tissue-specific methylation markers that agreed with clinical scores. Strikingly, transplanted patients with no clinical symptoms had abnormally high levels of tissue-specific markers, suggesting hidden tissue turnover even in the absence of evident clinical pathology. An integrative model taking into account total cfDNA concentration, monocyte/macrophage cfDNA levels and alanine transaminase was able to correctly identify GVHD with a specificity of 86% and precision of 89% (AUC of 0.8). CONCLUSION. cfDNA markers can be used for the detection of cGVHD, opening a window into underlying tissue dynamics in patients that receive allogeneic stem cell transplants. FUNDING. This work was supported by grants from the Ernest and Bonnie Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation, the Waldholtz/Pakula family, the Robert M. and Marilyn Sternberg Family Charitable Foundation and the Helmsley Charitable Trust (to YD)., Introduction Hematopoietic stem cell transplantation (HCT) is an essential and often the sole curative treatment strategy for high risk hematologic malignancies (1). Graft-versus-host disease (GVHD), the foremost complication of allogeneic [...]

A simple predictive analytical model for induction times in reverse osmosis (RO), both with and without an antiscalant (AS), has been developed based on the fundamental principles of mass and momentum balance. The simplicity of the model arises from the very low Reynolds number in the vicinity of the cluster surface, enabling the use and derivation of exact equations. The main assumption of the induction time without AS, t0A, is that the net growth of the cluster size results from the difference between adhesion and shear forces. With AS, the induction time, tA, is extended due to the competition between the AS and the scaling molecules on the cluster’s surface ligands. The model was validated by fitting it to six independent datasets from experiments conducted with spiral-wound and tubular RO membranes under various operational conditions, resulting in an average difference of 8.0% (t0A) and 8.7% (tA) between predicted and experimental induction times. It was found that t0A is governed by three dimensionless parameters: supersaturation ratio (Sa), shear (Ku), and scalant saturation (κ). tA increases with t0A and the AS concentration.

In the face of the SARS-CoV-2 pandemic, characterized by the virus’s rapid mutation rates, developing timely and targeted therapeutic and diagnostic interventions presents a significant challenge. This study utilizes bioinformatic analyses to pinpoint conserved genomic regions within SARS-CoV-2, offering a strategic advantage in the fight against this and future pathogens. Our approach has enabled the creation of a diagnostic assay that is not only rapid, reliable, and cost-effective but also possesses a remarkable capacity to detect a wide array of current and prospective variants with unmatched precision. The significance of our findings lies in the demonstration that focusing on these conserved genomic sequences can significantly enhance our preparedness for and response to emerging infectious diseases. By providing a blueprint for the development of versatile diagnostic tools and therapeutics, this research paves the way for a more effective global pandemic response strategy.

Reverse osmosis is used as a tertiary treatment for wastewater reclamation. However, sustainable management of the concentrate (ROC) is challenging, due to the need for treatment and/or disposal. The objective of this research was to investigate the efficiency of homogeneous and heterogeneous Fenton-like oxidation processes in removing propoxur (PR), a micro-pollutant compound, from synthetic ROC solution in a submerged ceramic membrane reactor operated in a continuous mode. A freshly prepared amorphous heterogeneous catalyst was synthesized and characterized, revealing a layered porous structure of 5–16 nm nanoparticles that formed aggregates (33–49 μm) known as ferrihydrite (Fh). The membrane exhibited a rejection of >99.6% for Fh. The homogeneous catalysis (Fe3+) exhibited better catalytic activity than the Fh in terms of PR removal efficiencies. However, by increasing the H2O2 and Fh concentrations at a constant molar ratio, the PR oxidation efficiencies were equal to those catalyzed by the Fe3+. The ionic composition of the ROC solution had an inhibitory effect on the PR oxidation, whereas increased residence time improved it up to 87% at a residence time of 88 min. Overall, the study highlights the potential of heterogeneous Fenton-like processes catalyzed by Fh in a continuous mode of operation. HIGHLIGHTS Propoxur removal from ROC solution in a submerged ceramic membrane reactor was studied.; Homogeneous (Fe3+) and heterogeneous (ferrihydrite) Fenton-like oxidation were compared.; While Fe3+ exhibited better catalytic activity than Fh, increased H2O2 and Fh concentrations led to similar PR removal as Fe3+.; Maximal PR removal of 87% was obtained at a residence time of 88 min.; The composition of the ROC inhibited the PR oxidation.;

Abstract Background Atopic dermatitis (AD) is one of the most common inflammatory skin diseases. It is associated with significant itch and impaired quality of life. Systemic treatments are efficient but associated with side effects. Novel topical treatments with a favourable safety profile are needed. SNG100 is a novel composition of hydrocortisone 1% in a cream base comprising sulphated polysaccharide (SPS; extracted from in‐house cultivated Porphyridium Cruentum unicellular algae), a well‐known hydrating, moisturising and a skin barrier repairing agent. Objectives To assess the safety, usability and efficacy of SNG100 cream in patients aged ≥6 years with moderate AD. Methods In this proof of concept phase I, double‐blind, randomised trial, participants received one of three treatments for 14 days: SNG100 twice daily (BID), hydrocortisone 1% BID or mometasone furoate once daily (QD). The primary endpoint was the safety and tolerability of SNG100 cream compared to hydrocortisone 1% and mometasone furoate. The secondary endpoint was the subject's usability of SNG100. Exploratory efficacy endpoints included percent change from baseline in SCOring AD (SCORAD), Eczema Area and Severity Index, Patient‐Oriented Eczema Measure, Dermatology Life Quality Index, pruritus Numerical Rating Score (NRS), peak pruritus‐NRS and Investigator's Global Assessment. Subjects were also followed up without any treatment for additional 14 days. Results Overall, 66 participants were screened, and 60 patients were randomised. SNG100 demonstrated a high safety profile, similar to marketed products hydrocortisone 1% and mometasone furoate 0.1%, with no unanticipated drug safety related events. SNG100 and mometasone furoate 0.1% cream achieved almost similar and statistically significant greater percentage reductions from baseline in SCORAD as compared to hydrocortisone 1% cream. SNG100 demonstrated significant improvement in NRS as compared to hydrocortisone 1% cream. Remarkably, SNG100 led to a lasting effect with only 29.4% of subjects returning to IGA3 during the follow‐up period compared to 50% and 38.9% in the hydrocortisone 1% and in mometasone furoate treatment arms, respectively. Conclusions Topical SNG100 is an effective, safe, and well‐tolerated innovative treatment for moderate AD. Trial registration number: NCT04615962 (Topical Cream SNG100 for Treatment in Moderate AD Subjects).

We present a methodology for a high-throughput screening (HTS) of transcription factor libraries, based on bacterial cells and GFP fluorescence. The method is demonstrated on the Escherichia coli LysR-type transcriptional regulator YhaJ, a key element in 2,4-dinitrotuluene (DNT) detection by bacterial explosives’ sensor strains. Enhancing the performance characteristics of the YhaJ transcription factor is essential for future standoff detection of buried landmines. However, conventional directed evolution methods for modifying YhaJ are limited in scope, due to the vast sequence space and the absence of efficient screening methods to select optimal transcription factor mutants. To overcome this limitation, we have constructed a focused saturation library of ca. 6.4 × 107 yhaJ variants, and have screened over 70 % of its sequence space using fluorescence-activated cell sorting (FACS). Through this screening process, we have identified YhaJ mutants exhibiting superior fluorescence responses to DNT, which were then effectively transformed into a bioluminescence-based DNT detection system. The best modified DNT reporter strain demonstrated a 7-fold lower DNT detection threshold, a 45-fold increased signal intensity, and a 40 % shorter response time compared to the parental bioreporter. The FACS-based HTS approach presented here may hold a potential for future molecular enhancement of other sensing and catalytic bioreactions.

microRNAs (miRNAs) are small non-coding RNAs (sncRNAs) that play an important role in the life cycle of human viruses. We sought to characterize human immunodeficiency virus 1 (HIV-1)-encoded miRNAs and determine their role in viral replication. Initially, a bioinformatic analysis was used to predict HIV-1-encoded miRNAs. Next, a representative number of these predicted sequences were verified using a miRNA microarray chip, reverse transcription PCR (RT-PCR), and the deep sequencing of RNA extracted from HIV-1-infected cells. Eight HIV-1-encoded sncRNA sequences conforming to the criteria that define miRNAs were identified in HIV-1-infected immortalized T cells and human primary CD4+ lymphocytes; five of the eight sequences have not been previously reported. Deep sequencing validated the presence of these virus-encoded miRNA sequences and uncovered large numbers of atypical sncRNA sequences, lacking characteristics of conventional miRNAs. We named these sequences small RNAs (smRNAs). The overexpression of four candidate HIV-1-encoded miRNAs and silencing of two smRNAs significantly increased HIV-1 viral replication. Our study uncovered novel HIV-1-encoded sncRNAs that, upon deregulated expression, alter viral titers in HIV-1-infected cells, suggesting that miRNAs and smRNAs play an important role in regulating viral replication. Future studies may reveal the function of HIV-1-encoded sncRNAs and their possible implications for diagnosis and treatment.

Nitrate pollution is a global problem as it affects both the environment and human health. The objective of this research was to study the effect of electrode configuration on the electro-reduction of nitrate. Coaxial cylindrical (inner rod and outer tube copper cathodes) and vertical plate parallel copper cathodes paired with Ti/RuO2-IrO2 (rod, tube, and plate) configurations were studied under various current densities and initial nitrate concentrations. The efficiency of each configuration was determined based on the removal efficiency of nitrate, specific energy consumption, mass transfer coefficients, and first order rate constants. Additionally, the overall systems’ resistance and geometric factors are discussed. It was found that the performances of the inner rod and outer tube copper cathodes were similar. The vertical plate parallel configuration was superior to the coaxial cylindrical electrode setup, as evident from a higher maximum nitrate removal of 74 and 56% at a current density of 7 mA/cm2 and lower energy consumption of 46.7 × 10−3 and 54.3 × 10−3 kWh/mmol NO3− at 36.4 mA/cm2, respectively. In addition, the mass transfer coefficients and first order rate constants were higher in all conditions tested for the vertical plate parallel configuration. HIGHLIGHTS Copper cathode was paired with Ti/RuO2-IrO2 anode for nitrate electro-reduction.; Performance of coaxial cylindrical and vertical plate configurations were compared.; The resistance, geometric factor and mass transfer coefficients were determined.; The effects of current density and initial nitrate concentrations were studied.; The coaxial cylindrical electrode setup was outperformed by the vertical plate parallel configuration.;