M2WISH: An easy and efficient protocol for whole-mount mRNA in situ hybridization that allows 3D cell resolution of gene expression in Arabidopsis thaliana.

Gene expression analysis is essential for understanding the mechanisms involved in plant development. Here, we developed M2WISH, a protocol based on MicroWave treatment for Wholemount mRNA In Situ Hybridization in Arabidopsis. By permeabilizing tissues without damaging cellular organization this protocol results in high and homogeneous hybridization yields that enable systematic analysis of gene expression dynamics. Moreover, when combined with cellular histochemical staining, M2WISH successfully provides a cellular resolution of gene expression. Thus, we demonstrate the robustness of M2WISH with 10 genes on roots, aerial meristems, leaves, and embryos in the seed. We applied M2WISH to study the spatial dynamics of WUSCHEL (WUS) and CLAVATA3 (CLV3) expression during in vitro meristematic conversion of roots into shoot apical meristems. Thus, we showed that shoot apical meristems could arise from two different types of root structures that differed by their CLV3 gene expression patterns. We constructed 3D cellular representations of WUS and CLV3 gene co-expression pattern and stressed the variability inherent to meristem conversion. Thus, this protocol generates a large amount of data on the localization of gene expression, which can be used to model complex systems.

Unveiling Dark Forces with Measurements of the Large Scale Structure of the Universe.

Cosmology offers opportunities to test dark matter independently of its interactions with the standard model. We study the imprints of long-range forces acting solely in the dark sector on the distribution of galaxies, the so-called large scale structure (LSS). We derive the strongest constraint on such forces from a combination of Planck and BOSS data. Along the way we consistently develop, for the first time, the effective field theory of LSS in the presence of new dynamics in the dark sector. We forecast that future surveys will improve the current bound by an order of magnitude.

Chemokine signaling via the CXCR2 receptor reinforces senescence.

Cells enter senescence, a state of stable proliferative arrest, in response to a variety of cellular stresses, including telomere erosion, DNA damage, and oncogenic signaling, which acts as a barrier against malignant transformation in vivo. To identify genes controlling senescence, we conducted an unbiased screen for small hairpin RNAs that extend the life span of primary human fibroblasts. Here, we report that knocking down the chemokine receptor CXCR2 (IL8RB) alleviates both replicative and oncogene-induced senescence (OIS) and diminishes the DNA-damage response. Conversely, ectopic expression of CXCR2 results in premature senescence via a p53-dependent mechanism. Cells undergoing OIS secrete multiple CXCR2-binding chemokines in a program that is regulated by the NF-kappaB and C/EBPbeta transcription factors and coordinately induce CXCR2 expression. CXCR2 upregulation is also observed in preneoplastic lesions in vivo. These results suggest that senescent cells activate a self-amplifying secretory network in which CXCR2-binding chemokines reinforce growth arrest.

Genome-wide identification of miRNAs and target regulatory network in the invasive ectoparasitic mite Varroa destructor.

Varroa destructor is an ectoparasite mite that attacks bees leading to colony disorders worldwide. microRNAs (miRNAs) are key molecules used by eukaryotes to post-transcriptional control of gene expression. Nevertheless, still lack information aboutV. destructor miRNAs and its regulatory networks. Here, we used an integrative strategy to characterize the miRNAs in the V. destructor mite. We identified 310 precursors that give rise to 500 mature miRNAs, which 257 are likely mite-specific elements. miRNAs showed canonical length ranging between 18 and 25 nucleotides and 5' uracil preference. Top 10 elements concentrated over 80% of total miRNA expression, with bantam alone representing ~50%. We also detected non-templated bases in precursor-derived small RNAs, indicative of miRNA post-transcriptional regulatory mechanisms. Finally, we note that conserved miRNAs control similar processes in different organisms, suggesting a conservative role. Altogether, our findings contribute to the better understanding of the mite biology that can assist future studies on varroosis control.

Anterior tibial tendon transfer in idiopathic clubfoot: does the outcome differ with the initial treatment? Proposed classification to surgical indication.

PURPOSE: To establish parameters and propose a radiographic classification for foot adduction/supination deformities to assist the indication of Garceau procedure. Secondly, to investigate whether the outcome of Garceau surgery depends on the initial treatment used, peritalar release, or the Ponseti method, and verify the maintenance of correction until skeletal maturity. METHODS: Prospective cohort study, with follow-up evaluations in 2009 and 2019. Fifty-three consecutive patients (71 feet) with idiopathic congenital clubfoot (ICCF) were divided into two groups according to the initial treatment used: peritalar release (group I) or Ponseti method (group II). All patients underwent Garceau procedure. The patients were evaluated clinically and radiographically using the American Orthopedic Foot Association (AOFAS) score for ankle and hindfoot. A radiographic classification is proposed based on the adduction by talo-first metatarsal angle on dorsoplantar view and supination by the elevation of the head of the first metatarsal on lateral view, both weight-bearing. RESULTS: In the first clinical evaluation, the mean score on the AOFAS was 87 points in group I and 86 points in group II. In the second evaluation, group I had mean AOFAS of 92 points and group II of 94 points. No statistical differences were found between the two groups, neither between the first and the second evaluation. In the radiographic evaluation, the adduction deformity obtained an average correction of 4° in group I and 3.6° in group II; in supination deformity, the average depression of the first metatarsal head in relation to the ground was 6.7 mm in group I and 7.5 mm in group II, with no statistical difference between the two evaluations, indicating the correction was maintained until skeletal maturity. CONCLUSION: Garceau transfer is capable of correcting residual deformities in adduction/supination of mild and moderate degrees, regardless of the initial treatment, and maintaining the correction until skeletal maturity. The proposed radiographic classification might help to objectively indicate the surgical procedure.

Open issues in management of carbon monoxide poisoning in pregnancy: practical suggestions.

Carbon monoxide (CO) poisoning during pregnancy may cause deleterious effects to the fetus. Hyperbaric oxygen therapy (HBO) in pregnancy is proven to be safe and it is considered to be beneficial, reducing the severity of the fetal injuries. However, a number of issues are still to be discussed, among them the question of the carboxyhemoglobin (COHb) levels that trigger HBO therapy in pregnant CO poisoned patients. In this letter we report some practical suggestions for organizations wishing to develop their own protocols.

Impact of COVID-19 pandemic on ST-elevation myocardial infarction in a non-COVID-19 epicenter.

OBJECTIVES: We sought to study the impact of COVID-19 pandemic on the presentation delay, severity, patterns of care, and reasons for delay among patients with ST-elevation myocardial infarction (STEMI) in a non-hot-spot region. BACKGROUND: COVID-19 pandemic has significantly reduced the activations for STEMI in epicenters like Spain. METHODS: From January 1, 2020, to April 15, 2020, 143 STEMIs were identified across our integrated 18-hospital system. Pre- and post-COVID-19 cohorts were based on March 23rd, 2020, whenstay-at-home orders were initiated in Ohio. We used presenting heart rate, blood pressure, troponin, new Q-wave, and left ventricle ejection fraction (LVEF) to assess severity. Duration of intensive care unit stay, total length of stay, door-to-balloon (D2B) time, and radial versus femoral access were used to assess patterns of care. RESULTS: Post-COVID-19 presentation was associated with a lower admission LVEF (45 vs. 50%, p = .015), new Q-wave, and higher initial troponin; however, these did not reach statistical significance. Among post-COVID-19 patients, those with >12-hr delay in presentation 31(%) had a longer average D2B time (88 vs. 53 min, p = .033) and higher peak troponin (58 vs. 8.5 ng/ml, p = .03). Of these, 27% avoided the hospital due to fear of COVID-19, 18% believed symptoms were COVID-19 related, and 9% did not want to burden the hospital during the pandemic. CONCLUSIONS: COVID-19 has remarkably affected STEMI presentation and care. Patients' fear and confusion about symptoms are integral parts of this emerging public health crisis.

Intracardiac echocardiography for guidance of transcatheter left atrial appendage occlusion: An expert consensus document.

Transcatheter left atrial appendage occlusion (LAAO) is an increasingly used alternative to oral anticoagulation in selected patients with atrial fibrillation. Intraprocedural imaging is a crucial for a successful intervention, with transesophageal echocardiography (TEE) as the current gold standard. Since some important limitations may affect TEE use, intracardiac echocardiography (ICE) is increasingly used as an alternative to TEE for guiding LAAO. The lack of a standardized imaging protocol has slowed the adoption of ICE into clinical practice. On the basis of current research and expert consensus, this paper provides a protocol for ICE support of left atrial appendage occlusion.

Flashing in-curb LEDs and beacons at unsignalized crosswalks and driver's visual attention to pedestrians during nighttime.

Driver's visual attention (eye movements) and driving behaviour (kinematic data) were assessed when approaching an experimental crosswalk that included flashing white in-curb LED strips, flashing orange beacons, backlit 'Yield here to pedestrians' vertical signs, and enhanced lighting when a staged pedestrian attempted to cross. An experimental condition in which all devices were active was compared with a control condition in which only enhanced lighting and backlit vertical signs were active. The results showed a significant increase of motorists' yielding compliance, distance of first fixation to the pedestrian area, standard deviation for horizontal eye movements in the experimental condition. The introduction of flashing in-curb LED strips and flashing orange beacons proved to be very effective in increasing the night-time safety of the pedestrian crossing. Practitioner summary: The study investigated the effects of flashing in-curb LED strips and beacons on driver's visual attention (eye movements) and speed when approaching a crosswalk during night-time. The results showed that the combination of these flashing devices significantly increased yielding compliance and the distance of pedestrian detection. Abbreviations: ANOVA: analysis of variance; EU: European Union; HAWK: high intensity activated crosswalk system; LED: light-emitting diode; PHB: pedestrian hybrid beacons; ROI: region of interest; RRFB: rectangular rapid flashing beacons; UK: United Kingdom; US: United States.

Direct conversion of root primordium into shoot meristem relies on timing of stem cell niche development.

To understand how the identity of an organ can be switched, we studied the transformation of lateral root primordia (LRP) into shoot meristems in Arabidopsis root segments. In this system, the cytokinin-induced conversion does not involve the formation of callus-like structures. Detailed analysis showed that the conversion sequence starts with a mitotic pause and is concomitant with the differential expression of regulators of root and shoot development. The conversion requires the presence of apical stem cells, and only LRP at stages VI or VII can be switched. It is engaged as soon as cell divisions resume because their position and orientation differ in the converting organ compared with the undisturbed emerging LRP. By alternating auxin and cytokinin treatments, we showed that the root and shoot organogenetic programs are remarkably plastic, as the status of the same plant stem cell niche can be reversed repeatedly within a set developmental window. Thus, the networks at play in the meristem of a root can morph in the span of a couple of cell division cycles into those of a shoot, and back, through transdifferentiation.

Visual Tension.

Although tension perception is well investigated in the music domain, its determinants in visual displays are still largely unexplored. Furthermore, the distinctive role of tension and arousal in affect theory is still debated. The study aimed to assess how geometrical and graphical transformations of basic visual shapes can affect perceived tension and arousal. The geometrical transformations were angle amplitude, rotation, position within a frame, symmetry, verticality, angularity, size, and regularity in spacing, while the graphical transformation regarded contrast. The sample included 122 participants. Perceived tension was significantly higher in angles with small amplitude, squares that were slightly rotated and not in the upright position, the upper and right areas within a rectangle, angular shapes, high-contrasted graphical transitions, asymmetrical shapes, vertical shapes, and dot patterns with irregular spacing. Overall, there was a moderate correlation between perception of tension and perception of arousal, although in some specific features, tension exhibited a dissociation from arousal, suggesting a distinctive role of tension in affect theory.

Productivity and nutritional value of BRS capiaçu grass (Pennisetum purpureum) managed at four regrowth ages in a semiarid region.

The objective of this study was to evaluate the productivity, productive efficiency, and nutritional value of the elephant grass cultivar BRS capiaçu (Pennisetum purpureum Schum.), managed at four regrowth ages during winter in the semiarid region of northern Minas Gerais, Brazil. A completely randomized design with the elephant grass cultivar BRS capiaçu was submitted to four cut intervals (30, 60, 90, and 120 days) in the winter with ten replications, for a total of 40 plots, each with a useful area of 6 × 5 m. There was a linear increase of 76.25% (P < 0.01) in the height of BRS capiaçu grass when cut from 30 to 120 days. Green matter production (P < 0.01) and dry matter production (P < 0.01) increased daily by 1081 kg/ha and 237 kg/ha, respectively. The annual dry matter production was 72 t/ha. Efficiency in water use changed (P < 0.01) from 7.91 kg of dry matter (DM)/mm at 30 days to 57.59 kg of DM/mm at 120 days of regrowth. There was a reduction in the ash content (P < 0.01), crude protein (P < 0.01), and the total digestible nutrient content (P < 0.01) with the increase in the age of the cut. The readily soluble fraction of DM (fraction A, P < 0.01), degradation rate "c" of insoluble fraction "B" (P = 0.01), potential degradability (PD; P < 0.01), and degradability (ED; P < 0.01) decreased linearly as the regrowth age increased. Harvesting is recommended at 90 days of regrowth during the winter season in this semiarid region.

Intracardiac echography for left atrial appendage closure: A step-by-step tutorial.

OBJECTIVES: We sought to provide a practical educational tool for the utilization of intracardiac echography (ICE) in the left atrium for the percutaneous closure of the left atrial appendage (LAA). BACKGROUND: Although transesophageal echocardiography (TEE) is the gold-standard imaging technique for LAA closure, ICE is stepping in to support noncoronary cardiology interventions by improving workflow and case turnover and may be more adequate for frail patients with significant and multiple comorbidities. METHODS: This article discusses the utility of ICE for LAA closure, its advantages compared to TEE, contraindications to TEE use and offers an extensive illustration of the main steps of the procedure. CONCLUSIONS: The use of ICE in the left atrium allows a feasible guidance of all steps of the percutaneous closure of LAA.

Impact of residual coronary atherosclerosis on transfemoral transcatheter aortic valve replacement.

OBJECTIVES: This study reports on the clinical effects of complete vs incompletely revascularized coronary artery disease on transcatheter aortic valve replacement (TAVR). BACKGROUND: There is a high prevalence of active coronary artery disease (CAD) in patients undergoing TAVR but preemptive revascularization remains controversial. METHODS: Patients were categorized into three cohorts: complete revascularization (CR), incomplete revascularization of a major epicardial artery (IR Major), and incomplete revascularization of a minor epicardial artery only (IR Minor). When feasible, SYNTAX scoring was performed for exploratory analysis. Analyses were performed using Cox proportional hazard models and Kaplan-Meier method. RESULTS: A total of 323 patients with active CAD were included. Adjusted outcomes showed that patients with IR Major had increased incidence of acute myocardial infarction (AMI) or revascularization compared with those in the CR cohort (HR 3.72, P = 0.048). No difference was noted in all-cause mortality or all-cause readmission rates. Exploratory secondary analysis with residual SYNTAX scores showed a significant interaction between disease burden and AMI/revascularization, as well as all-cause readmission. All-cause mortality remained unaffected based on residual SYNTAX scores. CONCLUSIONS: This is a retrospective single-center study reporting on pre-TAVR revascularization outcomes in patients with active CAD. In this analysis, we found that patients undergoing TAVR benefited from achieving complete revascularization to abate future incidence of AMI/revascularization. Despite this finding, all-cause mortality remained unaffected. Future efforts should focus on the role of functional assessment of the coronaries, as well as the long-term effects of complete revascularization in a larger patient cohort.

Comparison of intracardiac and transoesophageal echocardiography for guidance of percutaneous left atrial appendage occlusion: A meta-analysis.

AIMS: We performed a meta-analysis to compare the safety and efficacy of intracardiac echocardiography (ICE) vs transoesophageal echocardiography (TOE) for the guidance of percutaneous left atrial appendage occlusion (LAAO). METHODS AND RESULTS: A comprehensive search of the literature was performed, from inception to December 2018 using MEDLINE, Cochrane Library, and Google Scholar electronic databases, and references from articles. Publications comparing ICE and TOE for the guidance of percutaneous LAAO were included in the analysis. The co-primary endpoints were procedural success and procedure-related complications. Secondary endpoints included procedural and fluoroscopy times and the volume of contrast media used. Five studies were included in the analysis, providing a total of 1122 patients. Procedural success was similar with both techniques (pooled odd ratio [OR] 1.46, 95% confidence interval [CI] 0.77-2.75, P = 0.24, I2  = 0%), with a trend for less procedure-related complications in the ICE group (pooled OR 0.57, 95% CI 0.31-1.05, P = 0.07, I2  = 0%). Neither procedural (mean difference -1.64 minutes, 95% CI -16.42 to 13.14, P = 0.83, I2  = 96%) nor fluoroscopy times (mean difference + 1.29 minutes, 95% CI -1.38 to 3.97, P = 0.34, I2  = 76%) varied significantly between groups. There was a reduction in the volume of contrast media used in the ICE group (mean difference -9.71 mL, 95% CI -14.61 to -4.81, P > 0.01, I2  = 0%). CONCLUSION: Intracardiac echocardiography is a safe and effective alternative to TOE for guidance of percutaneous LAAO.

Risk Calculator to Predict 30-Day Readmission After Coronary Artery Bypass: A Strategic Decision Support Tool.

BACKGROUND: Re-admission is an important source of patient dissatisfaction and increased hospital costs. A simple calculator to determine the probability of re-admission may help guide patient dismissal planning. METHODS: Using the national readmissions database (NRD), we identified admissions for isolated primary coronary artery bypass (CABG) and stratified them according to 30-day readmission. Including pre, intra and postoperative variables, we prepared a logistic regression model to determine the probability for re-admission. The model was tested for reliability with boot-strapping and 10-fold cross-validation. RESULTS: From 135,699 procedures, 19,355 were readmitted at least once within 30days of dismissal. Patients who were readmitted were older (67±10 vs 65 ± 10 years, p<0.01), females (32% vs 24%; p<0.01) and had a higher Elixhauser comorbidity score (1.5±1.4 vs 1.1±1.2; p<0.01). Our final model (c- statistic=0.65) consisted of 16 pre and three postoperative factors. End-stage renal disease (OR 1.79 [1.57-2.04]) and length of stay>9days (OR 1.60 [1.52-1.68]) were most prominent indicators for readmission. Compared to Medicaid beneficiaries, those with private insurance (OR 0.62 [0.57-0.68]) and Medicare (OR 0.85 [0.79-0.92]) coverage were less likely to be readmitted. CONCLUSIONS: Our simple 30-days CABG readmission calculator can be used as a strategic tool to help reduce readmissions after coronary artery bypass surgery.

Safety of shorter length of hospital stay for patients undergoing minimalist transcatheter aortic valve replacement.

OBJECTIVE: Determine the feasibility and predictors of early discharge after minimalist transcatheter aortic valve replacement (TAVR). BACKGROUND: Duration of hospitalization has a direct impact on overall cost of care, but the clinical impact of length of stay (LOS) in patients undergoing minimalist TAVR remains unclear. METHODS: We studied 268 patients who underwent minimalist TAVR. Short LOS (sLOS) was defined as post-procedural LOS ≤ 3 days and observed in 163 patients. Prolonged LOS (pLOS) was observed in 105 patients. Propensity score matching based on 39 variables yielded 54 pairs of patients in each group. We analyzed 30-day mortality, 30-day re-hospitalization and long-term survival data. Multivariate regression models were used to define predictors of sLOS. RESULTS: Thirty-day mortality was 0% versus 5.5% in the sLOS and pLOS groups, respectively (P = 0.08). Incidence of re-hospitalization was higher in pLOS (13% vs. 3.7%). sLOS was associated with lower odds ratio of minor vascular complication (OR 0.1 [95% CI: 0.01, 0.75], P = 0.05), any bleeding (OR 0.35 [95% CI: 0.14, 0.87], P = 0.02), blood transfusion (OR 0.27 [95% CI: 0.08, 0.81], P = 0.02), and new pacemaker implantation (OR 0.23 [95% CI: 0.1, 0.53], P < 0.001). Discharge to home had a significantly higher odd ratio for sLOS (OR 8.67 [95% CI: 3.59, 23.11], P < 0.001). CONCLUSION: In appropriately selected patients, sLOS following minimalist TAVR approach in an experienced and high volume center is feasible and safe. Implementing such a strategy may reduce medical costs with the potential clinical benefit of early re-habilitation for the elderly TAVR population.

Thrombus aspiration in patients with ST-elevation myocardial infarction: results of a national registry of interventional cardiology.

BACKGROUND: We aimed to evaluate the impact of thrombus aspiration (TA) during primary percutaneous coronary intervention (P-PCI) in 'real-world' settings. METHODS: We performed a retrospective study, using data from the National Registry of Interventional Cardiology (RNCI 2006-2012, Portugal) with ST-elevation myocardial infarction (STEMI) patients treated with P-PCI. The primary outcome, in-hospital mortality, was analysed through adjusted odds ratio (aOR) and 95% confidence intervals (95%CI). RESULTS: We assessed data for 9458 STEMI patients that undergone P-PCI (35% treated with TA). The risk of in-hospital mortality with TA (aOR 0.93, 95%CI:0.54-1.60) was not significantly decreased. After matching patients through the propensity score, TA reduced significantly the risk of in-hospital mortality (OR 0.58, 95%CI:0.35-0.98; 3500 patients). CONCLUSIONS: The whole cohort data does not support the routine use of TA in P-PCI, but the results of the propensity-score matched cohort suggests that the use of selective TA may improve the short-term risks of STEMI.

Genetic Characterization of Some Neoponera (Hymenoptera: Formicidae) Populations Within the foetida Species Complex.

The foetida species complex comprises 13 Neotropical species in the ant genus Neoponera. Neoponera villosa Fabricius (1804) , Neoponera inversa Smith (1858), Neoponera bactronica Fernandes, Oliveira & Delabie (2013), and Neoponera curvinodis (Forel, 1899) have had an ambiguous taxonomic status for more than two decades. In southern Bahia, Brazil, these four species are frequently found in sympatry. Here we used Bayesian Inference and maximum likelihood analyses of COI and 16S mtDNA sequence data and conventional cytogenetic data together with observations on morphology to characterize sympatric populations of N. villosa, N. inversa, N. bactronica, and N. curvinodis. Our results showed marked differences in the karyotype of these ants. Both N. curvinodis and N. inversa have chromosome number of 2n = 30. Their chromosome composition, however, is distinct, which indicates that N. curvinodis is more closely related to N. bactronica. These four species clustered into three distinct groups. The close relationship between N. bactronica and N. curvinodis deserves further investigation since it has not been fully resolved here. Our results confirm that N. inversa, N. villosa, N. bactronica + N. curvinodis indeed represent four distinct taxa within the foetida species complex.

The role of peripheral vision in vertical road sign identification and discrimination.

The role of peripheral vision in road sign identification and discrimination was investigated in two studies. Peripheral vision plays an important role in road signs perception due to their lateral positioning. In the first study 20 participants identified road signs presented at five levels of horizontal eccentricity (1.1°-12.4°), and two levels of vertical eccentricity (0°-2.5°). In the second study road sign discrimination was tested in a same-different discrimination task. The first study showed that a vertical offset of 2.5° degraded proportion correct rate by 9%. Proportion correct rate decreased from 79% to 41% in the transition from 1.1° to 12.4° of horizontal offset. The second study showed an accurate discrimination for road signs presented within a horizontal offset of 6.4°. Road signs with angular shapes and prominent vertexes as triangular or cross signs were better identified in peripheral vision than signs with more compact shapes (circular signs). Practitioner summary: Vertical road signs, due to their lateral positioning, are often perceived in peripheral vision. Horizontal and vertical eccentricity negatively impacts the driver's ability to correctly identify and discriminate traffic signs. The use of singular shapes and a design with simple pictograms and large contrasting areas strongly facilitate road sign perception in peripheral vision.