Novel resistance to the Bymovirus BaMMV established by targeted mutagenesis of the PDIL5-1 susceptibility gene in barley.

The Potyviridae are the largest family of plant-pathogenic viruses. Members of this family are the soil-borne bymoviruses barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV), which, upon infection of young winter barley seedlings in autumn, can cause yield losses as high as 50%. Resistance breeding plays a major role in coping with these pathogens. However, some viral strains have overcome the most widely used resistance. Thus, there is a need for novel sources of resistance. In ancient landraces and wild relatives of cultivated barley, alleles of the susceptibility factor PROTEIN DISULFIDE ISOMERASE LIKE 5-1 (PDIL5-1) were identified to confer resistance to all known strains of BaYMV and BaMMV. Although the gene is highly conserved throughout all eukaryotes, barley is thus far the only species for which PDIL5-1-based virus resistance has been reported. Whereas introgression by crossing to the European winter barley breeding pool is tedious, time-consuming and additionally associated with unwanted linkage drag, the present study exemplifies an approach to targeted mutagenesis of two barley cultivars employing CRISPR-associated endonuclease technology to induce site-directed mutations similar to those described for PDIL5-1 alleles that render certain landraces resistant. Homozygous primary mutants were produced in winter barley, and transgene-free homozygous M2 mutants were produced in spring barley. A variety of mutants carrying novel PDIL5-1 alleles were mechanically inoculated with BaMMV, by which all frameshift mutations and certain in-frame mutations were demonstrated to confer resistance to this virus. Under greenhouse conditions, virus-resistant mutants showed no adverse effects in terms of growth and yield.

Antibiotic Cycling and Antibiotic Mixing: Which One Best Mitigates Antibiotic Resistance?

Can we exploit our burgeoning understanding of molecular evolution to slow the progress of drug resistance? One role of an infection clinician is exactly that: to foresee trajectories to resistance during antibiotic treatment and to hinder that evolutionary course. But can this be done at a hospital-wide scale? Clinicians and theoreticians tried to when they proposed two conflicting behavioral strategies that are expected to curb resistance evolution in the clinic, these are known as "antibiotic cycling" and "antibiotic mixing." However, the accumulated data from clinical trials, now approaching 4 million patient days of treatment, is too variable for cycling or mixing to be deemed successful. The former implements the restriction and prioritization of different antibiotics at different times in hospitals in a manner said to "cycle" between them. In antibiotic mixing, appropriate antibiotics are allocated to patients but randomly. Mixing results in no correlation, in time or across patients, in the drugs used for treatment which is why theorists saw this as an optimal behavioral strategy. So while cycling and mixing were proposed as ways of controlling evolution, we show there is good reason why clinical datasets cannot choose between them: by re-examining the theoretical literature we show prior support for the theoretical optimality of mixing was misplaced. Our analysis is consistent with a pattern emerging in data: neither cycling or mixing is a priori better than the other at mitigating selection for antibiotic resistance in the clinic. Key words: : antibiotic cycling, antibiotic mixing, optimal control, stochastic models.

Computerized versus hand-scored health literacy tools: a comparison of Simple Measure of Gobbledygook (SMOG) and Flesch-Kincaid in printed patient education materials.

OBJECTIVE: The research compared and contrasted hand-scoring and computerized methods of evaluating the grade level of patient education materials that are distributed at an academic medical center in east Tennessee and sought to determine if these materials adhered to the American Medical Association's (AMA's) recommended reading level of sixth grade. METHODS: Librarians at an academic medical center located in the heart of Appalachian Tennessee initiated the assessment of 150 of the most used printed patient education materials. Based on the Flesch-Kincaid (F-K) scoring rubric, 2 of the 150 documents were excluded from statistical comparisons due to the absence of text (images only). Researchers assessed the remaining 148 documents using the hand-scored Simple Measure of Gobbledygook (SMOG) method and the computerized F-K grade level method. For SMOG, 3 independent reviewers hand-scored each of the 150 documents. For F-K, documents were analyzed using Microsoft Word. Reading grade levels scores were entered into a database for statistical analysis. Inter-rater reliability was calculated using intra-class correlation coefficients (ICC). Paired t-tests were used to compare readability means. RESULTS: Acceptable inter-rater reliability was found for SMOG (ICC=0.95). For the 148 documents assessed, SMOG produced a significantly higher mean reading grade level (M=9.6, SD=1.3) than F-K (M=6.5, SD=1.3; p<0.001). Additionally, when using the SMOG method of assessment, 147 of the 148 documents (99.3%) scored above the AMA's recommended reading level of sixth grade. CONCLUSIONS: Computerized health literacy assessment tools, used by many national patient education material providers, might not be representative of the actual reading grade levels of patient education materials. This is problematic in regions like Appalachia because materials may not be comprehensible to the area's low-literacy patients. Medical librarians have the potential to advance their role in patient education to better serve their patient populations.

Foley catheter guide use during midurethral slings: does it make a difference?

INTRODUCTION: Our objective was to evaluate whether foley catheter guide use decreased the risk of cystotomy and urethrotomy during retropubic midurethral sling placement. MATERIALS AND METHODS: This retrospective cohort study included all women undergoing retropubic synthetic midurethral sling placement at a single academic institution between January 2011 and September 2012. Patients were divided into groups based on whether or not the foley catheter guide was used during surgery. The primary outcome was the incidence of cystotomy. RESULTS: A total of 310 patients underwent retropubic midurethral sling placement. The foley catheter guide was used in 76/310 cases (24.5%). The mean age was 57 ± 11 and mean body mass index was 28 ± 7. More patients in the no-guide group had preoperative urgency (70% versus 58%, p = 0.049), anterior prolapse (95% versus 78%, p < 0.0001), and concomitant prolapse surgery (65% versus 51%, p = 0.03). There was no difference in preoperative urgency urinary incontinence, medical comorbidities, previous surgical history, intraoperative time, blood loss, or postoperative voiding dysfunction rates between groups. Fourteen of the 310 patients (4.5%) had cystotomies: 1/76 (1.3%) in the foley catheter guide group and 13/234 (5.6%) in the no-guide group (p = 0.12). No patients had urethrotomies. On multiple logistic regression, there was no difference in the odds of cystotomy between groups after adjusting for previous prolapse and anti-incontinence surgery, concomitant prolapse repair, level of first assistant, and retropubic local anesthesia use (AOR = 0.2 [95% CI 0.02-1.7]). CONCLUSIONS: Foley catheter guide use did not decrease the risk of intraoperative lower urinary tract injury during retropubic midurethral sling placement. Larger prospective studies are needed to confirm this finding.

Development of the Biostatistics and Clinical Epidemiology Skills (BACES) assessment for medical residents.

BACKGROUND: Although biostatistics and clinical epidemiology are essential for comprehending medical evidence, research has shown consistently low and variable knowledge among postgraduate medical trainees. Simultaneously, there has been an increase in the complexity of statistical methods among top-tier medical journals. AIMS: To develop the Biostatics and Clinical Epidemiology Skills (BACES) assessment by (1) establishing content validity evidence of the BACES; (2) examining the model fit of the BACES items to an Item Response Theory (IRT) model; and (3) comparing IRT item estimates with those of traditional Classical Test Theory (CTT) indices. METHODS: Thirty multiple choice questions were written to focus on interpreting clinical epidemiological and statistical methods. Content validity was assessed through a four-person expert review. The instrument was administered to 150 residents across three academic medical centres in southern USA during the autumn of 2013. Data were fit to a two-parameter logistic IRT model and the item difficulty, discrimination and examinee ability values were compared with traditional CTT item statistics. RESULTS: 147 assessments were used for analysis (mean (SD) score 14.38 (3.38)). Twenty-six items, 13 devoted to statistics and 13 to clinical epidemiology, successfully fit a two-parameter logistic IRT model. These estimates also significantly correlated with their comparable CTT values. CONCLUSIONS: The strength of the BACES instrument was supported by (1) establishing content validity evidence; (2) fitting a sample of 147 residents' responses to an IRT model; and (3) correlating the IRT estimates with their CTT values, which makes it a flexible yet rigorous instrument for measuring biostatistical and clinical epidemiological knowledge.

Cold atmospheric plasma can effectively disinfect SARS-CoV-2 in the wastewater.

COVID-19 is currently pandemic and the detection of SARS-CoV-2 variants in wastewater is causing widespread concern. Herein, cold atmospheric plasma (CAP) is proposed as a novel wastewater disinfection technology that effectively inactivates SARS-CoV-2 transcription- and replication-competent virus-like particles, coronavirus GX_P2V, pseudotyped SARS-CoV-2 variants, and porcine epidemic diarrhoea virus in a large volume of water within 180 s (inhibition rate > 99%). Further, CAP disinfection did not adversely affect the viability of various human cell lines. It is identified that CAP produced peroxynitrite (ONOO-), ozone (O3), superoxide anion radicals (O2 -), and hydrogen peroxide (H2O2) as the major active substances for coronavirus disinfection. Investigation of the mechanism showed that active substances not only reacted with the coronavirus spike protein and affected its infectivity, but also destroyed the nucleocapsid protein and genome, thus affecting virus replication. This method provides an efficient and environmentally friendly strategy for the elimination of SARS-CoV-2 and other coronaviruses from wastewater.

A foundation model for clinical-grade computational pathology and rare cancers detection.

The analysis of histopathology images with artificial intelligence aims to enable clinical decision support systems and precision medicine. The success of such applications depends on the ability to model the diverse patterns observed in pathology images. To this end, we present Virchow, the largest foundation model for computational pathology to date. In addition to the evaluation of biomarker prediction and cell identification, we demonstrate that a large foundation model enables pan-cancer detection, achieving 0.95 specimen-level area under the (receiver operating characteristic) curve across nine common and seven rare cancers. Furthermore, we show that with less training data, the pan-cancer detector built on Virchow can achieve similar performance to tissue-specific clinical-grade models in production and outperform them on some rare variants of cancer. Virchow's performance gains highlight the value of a foundation model and open possibilities for many high-impact applications with limited amounts of labeled training data.

A specific fine-grained identification model for plasma-treated rice growth using multiscale shortcut convolutional neural network.

As an agricultural innovation, low-temperature plasma technology is an environmentally friendly green technology that increases crop quality and productivity. However, there is a lack of research on the identification of plasma-treated rice growth. Although traditional convolutional neural networks (CNN) can automatically share convolution kernels and extract features, the outputs are only suitable for entry-level categorization. Indeed, shortcuts from the bottom layers to fully connected layers can be established feasibly in order to utilize spatial and local information from the bottom layers, which contain small distinctions necessary for fine-grain identification. In this work, 5000 original images which contain the basic growth information of rice (including plasma treated rice and the control rice) at the tillering stage were collected. An efficient multiscale shortcut CNN (MSCNN) model utilizing key information and cross-layer features was proposed. The results show that MSCNN outperforms the mainstream models in terms of accuracy, recall, precision and F1 score with 92.64%, 90.87%, 92.88% and 92.69%, respectively. Finally, the ablation experiment, comparing the average precision of MSCNN with and without shortcuts, revealed that the MSCNN with three shortcuts achieved the best performance with the highest precision.

Avoiding falsely low creatinine concentrations measured in patients treated with N-acetylcysteine for acetaminophen intoxication using enzymo-amperometric method - An in vitro and in vivo study.

BACKGROUND: Circulating creatinine is a biomarker of paramount importance in clinical practice. In cases of acetaminophen (APAP) intoxication, the antidote, N-acetylcysteine (NAC), interferes with commonly used creatininase-peroxidase methods. This study aimed to assess whether creatininase-amperometric methods were affected in this context. METHODS: This study includes in vitro interference tests, involving four creatinine assays using NAC-spiked plasma pools and an in vivo retrospective study comparing creatininase-peroxidase and creatininase-amperometric measurements in patients presenting with NAC-treated APAP poisoning. RESULTS: Creatininase-peroxidase method was impacted by NAC interference in a clinically-significant manner at therapeutic NAC levels (basal value recovery of 80 % and 70 % for 500 and 1000 mg.L-1 of NAC, respectively), surpassing the desirable Reference Change Value (RCV%). Enzymo-amperometric methods were not impacted. Among patients, a mean bias of -45.2 ± 28.0 % was observed for the peroxidase detection method compared to the amperometric in those who received NAC prior plasma sampling and -2.7 ± 5.4 % in those who did not. CONCLUSIONS: Our findings indicate that enzymo-amperometric creatinine assays remain unaffected by NAC interference due to the absence of the peroxidase step in the analytical process. Therefore, these methods are suitable to prevent spurious hypocreatininemia in APAP intoxicated patients undergoing NAC therapy.

The earliest unambiguous Neanderthal engravings on cave walls: La Roche-Cotard, Loire Valley, France.

Here we report on Neanderthal engravings on a cave wall at La Roche-Cotard (LRC) in central France, made more than 57±3 thousand years ago. Following human occupation, the cave was completely sealed by cold-period sediments, which prevented access until its discovery in the 19th century and first excavation in the early 20th century. The timing of the closure of the cave is based on 50 optically stimulated luminescence ages derived from sediment collected inside and from around the cave. The anthropogenic origin of the spatially-structured, non-figurative marks found within the cave is confirmed using taphonomic, traceological and experimental evidence. Cave closure occurred significantly before the regional arrival of H. sapiens, and all artefacts from within the cave are typical Mousterian lithics; in Western Europe these are uniquely attributed to H. neanderthalensis. We conclude that the LRC engravings are unambiguous examples of Neanderthal abstract design.

ROS implication in a new antitumor strategy based on non-thermal plasma.

Non-thermal plasma (NTP) is generated by ionizing neutral gas molecules/atoms leading to a highly reactive gas at ambient temperature containing excited molecules, reactive species and generating transient electric fields. Given its potential to interact with tissue or cells without a significant temperature increase, NTP appears as a promising approach for the treatment of various diseases including cancer. The aim of our study was to evaluate the interest of NTP both in vitro and in vivo. To this end, we evaluated the antitumor activity of NTP in vitro on two human cancer cell lines (glioblastoma U87MG and colorectal carcinoma HCT-116). Our data showed that NTP generated a large amount of reactive oxygen species (ROS), leading to the formation of DNA damages. This resulted in a multiphase cell cycle arrest and a subsequent apoptosis induction. In addition, in vivo experiments on U87MG bearing mice showed that NTP induced a reduction of bioluminescence and tumor volume as compared to nontreated mice. An induction of apoptosis was also observed together with an accumulation of cells in S phase of the cell cycle suggesting an arrest of tumor proliferation. In conclusion, we demonstrated here that the potential of NTP to generate ROS renders this strategy particularly promising in the context of tumor treatment.

A van der Waals force-based adhesion study of stem cells exposed to cold atmospheric plasma jets.

Cold atmospheric plasma has established its effect on cell adhesion. Given the importance of cell adhesion in stem cells, the current study investigates the effect of plasma treatment on Human Bone Marrow Mesenchymal Stem Cells (HBMMSCs) adhesion by which the differentiation and fate of cells are determined. In this paper, adhesion modification is considered not only for cell- ECM (Extra cellular Matrix), but also between suspended cells, and enhanced adhesions were found in both circumstances. Regarding the previous works, the increase of the cell-ECM adhesion during the plasma therapy was mostly attributed to the enhancement of the production and activity of integrin proteins. Nevertheless, considering the importance of van der Waals forces at the cellular level, the effect of cold plasma on VDWFs and so its effect on adhesion is investigated in this work for the first time, to the best of our knowledge. For this purpose, employing the semi-empirical methods, the role of the plasma therapy on the VDWF between the cells has been studied at three levels; (a) plasma-induced dipole formation, (b) Hammaker coefficient modification of culture medium, and c) cell roughness modification. For suspended cell condition, we conclude and support that van der Waals forces (VDWFs) enhancement has a key role in cell adhesion processes. We believe that, the present work gives a new physical insight in studying the plasma therapy method at the cellular level.

Site-Directed Mutagenesis in Barley Using RNA-Guided Cas Endonucleases During Microspore-Derived Generation of Doubled Haploids.

In plant research and breeding, haploid technology is employed upon crossing, induced mutagenesis or genetic engineering to generate populations of meiotic recombinants that are themselves genetically fixed. Thanks to the speed and efficiency in producing true-breeding lines, haploid technology has become a major driver of modern crop improvement. In the present study, we used embryogenic pollen cultures of winter barley ( Hordeum vulgare ) for Cas9 endonuclease-mediated targeted mutagenesis in haploid cells, which facilitates the generation of homozygous primary mutant plants. To this end, microspores were extracted from immature anthers, induced to undergo cell proliferation and embryogenic development in vitro, and were then inoculated with Agrobacterium for the delivery of T-DNAs comprising expression units for Cas9 endonuclease and target gene-specific guide RNAs (gRNAs). Amongst the regenerated plantlets, mutants were identified by PCR amplification of the target regions followed by sequencing of the amplicons. This approach also enabled us to discriminate between homozygous and heterozygous or chimeric mutants. The heritability of induced mutations and their homozygous state were experimentally confirmed by progeny analyses. The major advantage of the method lies in the preferential production of genetically fixed primary mutants, which facilitates immediate phenotypic analyses and, relying on that, a particularly efficient preselection of valuable lines for detailed investigations using their progenies.

Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley.

The Eukaryotic Translation Initiation Factor 4E (EIF4E) is a well-known susceptibility factor for potyvirus infections in many plant species. The barley yellow mosaic virus disease, caused by the bymoviruses Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), can lead to yield losses of up to 50% in winter barley. In autumn, the roots of young barley plants are infected by the soil-borne plasmodiophoraceous parasite Polymyxa graminis L. that serves as viral vector. Upon viral establishment and systemic spreading into the upper parts of the plants, yellow mosaics occur as first symptoms on leaves. In the further course of plant development, the disease entails leaf necrosis and increased susceptibility to frost damage. Thanks to the rym4 and rym5 allelic variants of the HvEIF4E gene, more than two thirds of current European winter barley cultivars are resistant to BaYMV and BaMMV. However, several strains of BaYMV and BaMMV have already overcome rym4- and rym5-mediated resistance. Accordingly, new resistance-conferring alleles are needed for barley breeding. Therefore, we performed targeted mutagenesis of the EIF4E gene by Cas9 endonuclease in BaMMV/BaYMV-susceptible winter barley cv. "Igri". Small insertions were generated, resulting in a shift of the translational reading frame, thereby causing the loss-of-function of EIF4E. The mutations occurred in the homozygous state already in the primary mutants. Their progeny proved invariably homozygous and fully resistant to mechanical inoculation with BaMMV. EIF4E knockout plants showed normal growth habit and produced grains, yet exhibited a yield penalty.

The emerging potential of cold atmospheric plasma in skin biology.

The maintenance of skin integrity is crucial to ensure the physiological barrier against exogenous compounds, microorganisms and dehydration but also to fulfill social and aesthetic purposes. Besides the development of new actives intended to enter a formulation, innovative technologies based on physical principles have been proposed in the last years. Among them, Cold Atmospheric Plasma (CAP) technology, which already showed interesting results in dermatology, is currently being studied for its potential in skin treatments and cares. CAP bio-medical studies gather several different expertise ranging from physics to biology through chemistry and biochemistry, making this topic hard to pin. In this review we provide a broad survey of the interactions between CAP and skin. In the first section, we tried to give some fundamentals on skin structure and physiology, related to its essential functions, together with the main bases on cold plasma and its physicochemical properties. In the following parts we dissected and analyzed each CAP parameter to highlight the already known and the possible effects they can play on skin. This overview aims to get an idea of the potential of cold atmospheric plasma technology in skin biology for the future developments of dermo-cosmetic treatments, for example in aging prevention.

Cell Electropermeabilisation Enhancement by Non-Thermal-Plasma-Treated PBS.

The effectiveness of electrochemotherapy (ECT) in local eradication of tumours in human and veterinary medicine has been proven. ECT consists of increasing the uptake of cytotoxic drugs by means of pulsed electric fields (PEFs) that transiently permeabilise the cell membrane. Still, this tumour treatment includes some drawbacks that are linked to the characteristics of the intense electric pulses (EPs) used. Meanwhile, the emerging field of cancer therapies that are based on the application of non-thermal plasmas (NTP) has recently garnered interest because of their potentialities as rich sources of reactive species. In this work, we investigated the potential capabilities of the combined application of indirect NTP treatment and microsecond PEFs (µsPEFs) to outperform in vitro cell electropermeabilisation, the basis of ECT. Thus, phosphate-buffered saline (PBS) was plasma-treated (pPBS) and used afterwards to explore the effects of its combination with µsPEFs. Analysis of two different cell lines (DC-3F Chinese hamster lung fibroblasts and malignant B16-F10 murine melanoma cells), by flow cytometry, revealed that this combination resulted in significant increases of the level of cell membrane electropermeabilisation, even at very low electric field amplitude. The B16-F10 cells were more sensitive to the combined treatment than DC-3F cells. Importantly, the percentage of permeabilised cells reached values similar to those of cells exposed to classical electroporation field amplitude (1100 V/cm) when the cells were treated with pPBS before and after being exposed only to very low PEF amplitude (600 V/cm). Although the level of permeabilisation of the cells that are treated by the pPBS and the PEFs at 600 V/cm is lower than the level reached after the exposure to µsPEFs alone at 1100 V/cm, the combined treatment opens the possibility to reduce the amplitude of the EPs used in ECT, potentially allowing for a novel ECT with reduced side-effects.

Evaluating and Exploring Variations in Surgical Resident Emotional Intelligence and Burnout.

OBJECTIVE: Surgery resident burnout rates are on the rise, ranging from 50% to 69%. Burnout is associated with increased risk of error and poorer patient satisfaction. Emotional intelligence (EI) is defined as the capacity to be aware of, control, and express one's emotions, and to handle interpersonal relationships judiciously and empathetically. We seek to evaluate the correlation between EI and burnout temporally as a potential target for education. This may allow us to utilize objective measures to reduce burnout among our residents. DESIGN: A prospective study of general surgery residents at a single institution was performed via self-reporting assessments on personal demographics, the Maslach Burnout Inventory (MBI), and the Scale of Emotional Functioning: Health Service Provider at 3 separate time-points. SETTING: A medium sized academic medical center in Tennessee approved to graduate 6 chief residents per year. PARTICIPANTS: All general surgery residents PGY1 to PGY5 including preliminary residents were given the assessment tools and the option of participating. Research residents were excluded. RESULTS: A total of 86 assessments were completed, including 15 residents who completed all 3 assessments. Changes in the personal achievement (PA) portion of the MBI had the strongest correlation with temporal changes in EI with a Pearson correlation coefficient of 0.606 and 0.616 (p 0.017 and 0.015, respectively). Of the 3 subscales of the MBI, residents had moderate or severe emotional exhaustion for 62% of responses, 73% for depersonalization, and 37% for PA. All categories examined (emotional exhaustion, depersonalization, PA, and EI) saw improvement in scores across 3 administrations. CONCLUSIONS: Improvements in burnout and EI scores were seen across 3 administrations of surveys without any intervention. In the future, assessing PA may have the highest potential to evaluate burnout indirectly. Designing a curriculum for EI may assist with preventing burnout.

An assisted-living home architecture with integrated healthcare services for elderly people.

Since the population of elderly people grows absolutely and in relation to the overall population in the world, the improvement of the quality of life of elderly people at home is of a great importance. This can be achieved through the development of generic technologies for managing their domestic ambient environment consisting of medical sensors, entertainment equipment, home automation systems and white goods, increasing their autonomy and safety. In this context, the provision intelligent interactive healthcare services will improve their daily life and allowing at the same time the continuous monitoring of their health and their effective treatment. This work is supported by the INHOME Project EU IST-045061-STP, http://www.ist-inhome.eu.