Immunofluorescent Detection of Two Thymidine Analogues (CldU and IdU) in Primary Tissue

Accurate measurement of cell division is a fundamental challenge in experimental biology that becomes increasingly complex when slowly dividing cells are analyzed. Established methods to detect cell division include direct visualization by continuous microscopy in cell culture, dilution of vital dyes such as carboxyfluorescein di-aetate succinimidyl ester (CFSE), immuno-detection of mitogenic antigens such as ki67 or PCNA, and thymidine analogues. Thymidine analogues can be detected by a variety of methods including radio-detection for tritiated thymidine, immuno-detection for bromo-deoxyuridine (BrdU), chloro-deoxyuridine (CldU) and iodo-deoxyuridine (IdU), and chemical detection for ethinyl-deoxyuridine (EdU). We have derived a strategy to detect sequential incorporation of different thymidine analogues (CldU and IdU) into tissues of adult mice. Our method allows investigators to accurately quantify two successive rounds of cell division. By optimizing immunostaining protocols our approach can detect very low dose thymidine analogues administered via the drinking water, safe to administer to mice for prolonged periods of time. Consequently, our technique can be used to detect cell turnover in very long-lived tissues. Optimal immunofluoresent staining results can be achieved in multiple tissue types, including pancreas, skin, gut, liver, adrenal, testis, ovary, thyroid, lymph node, and brain. We have also applied this technique to identify oncogenic transformation within tissues. We have further applied this technique to determine if transit-amplifying cells contribute to growth or renewal of tissues. In this sense, sequential administration of thymidine analogues represents a novel approach for studying the origins and survival of cells involved in tissue homeostasis

BAT AGN Spectroscopic Survey – XIII. The nature of the most luminous obscured AGN in the low-redshift universe

We present a multiwavelength analysis of 28 of the most luminous low-redshift narrow-line, ultra-hard X-ray-selected active galactic nuclei (AGN) drawn from the 70-month Swift/BAT all-sky survey, with bolometric luminosities of log(L_(bol)/ergs^(-1)) ≳ 45.25⁠. The broad goal of our study is to determine whether these objects have any distinctive properties, potentially setting them aside from lower luminosity obscured AGN in the local Universe. Our analysis relies on the first data release of the BAT AGN Spectroscopic Survey (BASS/DR1) and on dedicated observations with the VLT, Palomar, and Keck observatories. We find that the vast majority of our sources agree with commonly used AGN selection criteria which are based on emission line ratios and on mid-infrared colours. Our AGN are pre-dominantly hosted in massive galaxies (9.8 ≲ log (M*/M⊙) ≲ 11.7); based on visual inspection of archival optical images, they appear to be mostly ellipticals. Otherwise, they do not have distinctive properties. Their radio luminosities, determined from publicly available survey data, show a large spread of almost four orders of magnitude – much broader than what is found for lower X-ray luminosity obscured AGN in BASS. Moreover, our sample shows no preferred combination of black hole masses (MBH) and/or Eddington ratio (λ_(Edd)), covering 7.5 ≲ log (M_(BH)/M⊙) ≲ 10.3 and 0.01 ≲ λ_(Edd) ≲ 1. Based on the distribution of our sources in the λ_(Edd)−N_H plane, we conclude that our sample is consistent with a scenario where the amount of obscuring material along the line of sight is determined by radiation pressure exerted by the AGN on the dusty circumnuclear gas

BAT AGN Spectroscopic Survey - XIII. The nature of the most luminous obscured AGN in the low-redshift universe

ISSN:0035-8711ISSN:1365-2966ISSN:1365-871

Probing the Structure and Evolution of BASS AGN through Eddington Ratios

We constrain the intrinsic Eddington ratio (\lamEdd ) distribution function for local AGN in bins of low and high obscuration (log NH <= 22 and 22 < log NH < 25), using the Swift-BAT 70-month/BASS DR2 survey. We interpret the fraction of obscured AGN in terms of circum-nuclear geometry and temporal evolution. Specifically, at low Eddington ratios (log lamEdd < -2), obscured AGN outnumber unobscured ones by a factor of ~4, reflecting the covering factor of the circum-nuclear material (0.8, or a torus opening angle of ~ 34 degrees). At high Eddington ratios (\log lamEdd > -1), the trend is reversed, with < 30% of AGN having log NH > 22, which we suggest is mainly due to the small fraction of time spent in a highly obscured state. Considering the Eddington ratio distribution function of narrow-line and broad-line AGN from our prior work, we see a qualitatively similar picture. To disentangle temporal and geometric effects at high lamEdd, we explore plausible clearing scenarios such that the time-weighted covering factors agree with the observed population ratio. We find that the low fraction of obscured AGN at high lamEdd is primarily due to the fact that the covering factor drops very rapidly, with more than half the time is spent with < 10% covering factor. We also find that nearly all obscured AGN at high-lamEdd exhibit some broad-lines. We suggest that this is because the height of the depleted torus falls below the height of the broad-line region, making the latter visible from all lines of sight.Comment: Accepted by ApJ

Ursolic Acid Increases Skeletal Muscle and Brown Fat and Decreases Diet-Induced Obesity, Glucose Intolerance and Fatty Liver Disease

Skeletal muscle Akt activity stimulates muscle growth and imparts resistance to obesity, glucose intolerance and fatty liver disease. We recently found that ursolic acid increases skeletal muscle Akt activity and stimulates muscle growth in non-obese mice. Here, we tested the hypothesis that ursolic acid might increase skeletal muscle Akt activity in a mouse model of diet-induced obesity. We studied mice that consumed a high fat diet lacking or containing ursolic acid. In skeletal muscle, ursolic acid increased Akt activity, as well as downstream mRNAs that promote glucose utilization (hexokinase-II), blood vessel recruitment (Vegfa) and autocrine/paracrine IGF-I signaling (Igf1). As a result, ursolic acid increased skeletal muscle mass, fast and slow muscle fiber size, grip strength and exercise capacity. Interestingly, ursolic acid also increased brown fat, a tissue that shares developmental origins with skeletal muscle. Consistent with increased skeletal muscle and brown fat, ursolic acid increased energy expenditure, leading to reduced obesity, improved glucose tolerance and decreased hepatic steatosis. These data support a model in which ursolic acid reduces obesity, glucose intolerance and fatty liver disease by increasing skeletal muscle and brown fat, and suggest ursolic acid as a potential therapeutic approach for obesity and obesity-related illness

Too Hot to Handle: An Evaluation of the Effect of Thermal Visual Representation on User Grasping Interaction in Virtual Reality

Influence of interaction fidelity and rendering quality on perceived user experience have been largely explored in Virtual Reality (VR). However, differences in interaction choices triggered by these rendering cues have not yet been explored. We present a study analysing the effect of thermal visual cues and contextual information on 50 participants' approach to grasp and move a virtual mug. This study comprises 3 different temperature cues (baseline empty, hot and cold) and 4 contextual representations; all embedded in a VR scenario. We evaluate 2 different hand representations (abstract and human) to assess grasp metrics. Results show temperature cues influenced grasp location, with the mug handle being predominantly grasped with a smaller grasp aperture for the hot condition, while the body and top were preferred for baseline and cold conditions

The Death and Rebirth of a Party System, Peru 1978-2001

This article evaluates structural, institutional, and actor-centered explanations of the collapse of the Peruvian party system around 1990 and its surprising partial recovery in 2001. It begins by describing the changes in the dependent variable, the emergence, collapse, and partial resurrection of the 1980s Peruvian party system. The next section examines the argument that the large size and rapid growth of the informal sector undermined the party system and led to its collapse. The author shows that the evidence does not support this argument. The article then examines changes in the electoral system. The author demonstrates that, contrary to theoretical expectations, the changes in the electoral system do not correlate with the observed changes in the party system. The final section shows that performance failure by political elites, including corruption in government, was more important than social cleavages or electoral institutions in the collapse and partial recovery of the party system.Yeshttps://us.sagepub.com/en-us/nam/manuscript-submission-guideline

Post–COVID-19 Conditions Among Children 90 Days After SARS-CoV-2 Infection

IMPORTANCE Little is known about the risk factors for, and the risk of, developing post-COVID-19 conditions (PCCs) among children. OBJECTIVES To estimate the proportion of SARS-CoV-2-positive children with PCCs 90 days after a positive test result, to compare this proportion with SARS-CoV-2-negative children, and to assess factors associated with PCCs. DESIGN, SETTING, AND PARTICIPANTS This prospective cohort study, conducted in 36 emergency departments (EDs) in 8 countries between March 7, 2020, and January 20, 2021, included 1884 SARS-CoV-2-positive children who completed 90-day follow-up; 1686 of these children were frequency matched by hospitalization status, country, and recruitment date with 1701 SARS-CoV-2-negative controls. EXPOSURE SARS-CoV-2 detected via nucleic acid testing. MAIN OUTCOMES AND MEASURES Post-COVID-19 conditions, defined as any persistent, new, or recurrent health problems reported in the 90-day follow-up survey. RESULTS Of 8642 enrolled children, 2368 (27.4%) were SARS-CoV-2 positive, among whom 2365 (99.9%) had index ED visit disposition data available; among the 1884 children (79.7%) who completed follow-up, the median age was 3 years (IQR, 0-10 years) and 994 (52.8%) were boys. A total of 110 SARS-CoV-2-positive children (5.8%; 95% CI, 4.8%-7.0%) reported PCCs, including 44 of 447 children (9.8%; 95% CI, 7.4%-13.0%) hospitalized during the acute illness and 66 of 1437 children (4.6%; 95% CI, 3.6%-5.8%) not hospitalized during the acute illness (difference. 5.3%; 95% CI, 2.5%-8.5%). Among SARS-CoV-2-positive children, the most common symptom was fatigue or weakness (21 [1.1%]). Characteristics associated with reporting at least 1 PCC at 90 days included being hospitalized 48 hours or more compared with no hospitalization (adjusted odds ratio [aOR], 2.67 [95% CI, 1.63-4.38]); having 4 or more symptoms reported at the index ED visit compared with 1 to 3 symptoms (4-6 symptoms: aOR, 2.35 [95% CI, 1.28-4.31]; >= 7 symptoms: aOR, 4.59 [95% CI, 2.50 8.44]); and being 14 years of age or older compared with younger than 1 year (aOR, 2.67 [95% CI, 1.43-4.99]). SARS-CoV-2-positive children were more likely to report PCCs at 90 days compared with those who tested negative, both among those who were not hospitalized (55 of 1295 [4.2%; 95% CI, 3.2%-5.5%] vs 35 of 1321[2.7%; 95% CI, 1.9%-3.7%]; difference, 1.6% [95% CI, 0.2%-3.0%]) and those who were hospitalized (40 of 391[10.2%; 95% CI, 7.4%-13.7%] vs 19 of 380 [5.0%; 95% CI, 3.0%-7.7%]; difference, 5.2% [95% CI, 1.5%-9.1%]). In addition, SARS-CoV-2 positivity was associated with reporting PCCs 90 days after the index ED visit (aOR, 1.63 [95% CI, 1.14-2.35]), specifically systemic health problems (eg, fatigue, weakness, fever; aOR, 2.44 [95% CI, 1.19-5.00]). CONCLUSIONS AND RELEVANCE In this cohort study, SARS-CoV-2 infection was associated with reporting PCCs at 90 days in children. Guidance and follow-up are particularly necessary for hospitalized children who have numerous acute symptoms and are older.This studywas supported by grants from the Canadian Institutes of Health Research (operating grant: COVID-19-clinical management); the Alberta Health Services-University of Calgary-Clinical Research Fund; the Alberta Children's Hospital Research Institute; the COVID-19 Research Accelerator Funding Track (CRAFT) Program at the University of California, Davis; and the Cincinnati Children's Hospital Medical Center Division of Emergency Medicine Small Grants Program. Dr Funk is supported by the University of Calgary Eyes-High PostDoctoral Research Fund. Dr Freedman is supported by the Alberta Children's Hospital Foundation Professorship in Child Health andWellness