The Mycoplasma hyorhinis genome displays differential chromatin accessibility.

Whilst the regulation of chromatin accessibility and its effect on gene expression have been well studied in eukaryotic species, the role of chromatin dynamics and 3D organisation in genome reduced bacteria remains poorly understood [1,2]. In this study we profiled the accessibility of the Mycoplasma hyorhinis genome, these data were collected fortuitously as part of an experiment where ATAC-Seq was conducted on mycoplasma, contaminated mammalian cells. We found a differential and highly reproducible chromatin accessibility landscape, with regions of increased accessibility corresponding to genes important for the bacteria's life cycle and infectivity. Furthermore, accessibility in general correlated with transcriptionally active genes as profiled by RNA-Seq, but peaks of high accessibility were also seen in non-coding and intergenic regions, which could contribute to the topological organisation of the genome. However, changes in transcription induced by starvation or application of the RNA polymerase inhibitor rifampicin did not themselves change the accessibility profile, which confirms that the differential accessibility is inherently a property of the genome, and not a consequence of its function. These results together show that differential chromatin accessibility is a key feature of the regulation of gene expression in bacteria.

The circadian clock component BMAL1 regulates SARS-CoV-2 entry and replication in lung epithelial cells.

The coronavirus disease 2019 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) coronavirus, is a global health issue with unprecedented challenges for public health. SARS-CoV-2 primarily infects cells of the respiratory tract via spike glycoprotein binding to angiotensin-converting enzyme (ACE2). Circadian rhythms coordinate an organism's response to its environment and can regulate host susceptibility to virus infection. We demonstrate that silencing the circadian regulator Bmal1 or treating lung epithelial cells with the REV-ERB agonist SR9009 reduces ACE2 expression and inhibits SARS-CoV-2 entry and replication. Importantly, treating infected cells with SR9009 limits SARS-CoV-2 replication and secretion of infectious particles, showing that post-entry steps in the viral life cycle are influenced by the circadian system. Transcriptome analysis revealed that Bmal1 silencing induced interferon-stimulated gene transcripts in Calu-3 lung epithelial cells, providing a mechanism for the circadian pathway to limit SARS-CoV-2 infection. Our study highlights alternative approaches to understand and improve therapeutic targeting of SARS-CoV-2.

Adenosine integrates light and sleep signalling for the regulation of circadian timing in mice.

The accumulation of adenosine is strongly correlated with the need for sleep and the detection of sleep pressure is antagonised by caffeine. Caffeine also affects the circadian timing system directly and independently of sleep physiology, but how caffeine mediates these effects upon the circadian clock is unclear. Here we identify an adenosine-based regulatory mechanism that allows sleep and circadian processes to interact for the optimisation of sleep/wake timing in mice. Adenosine encodes sleep history and this signal modulates circadian entrainment by light. Pharmacological and genetic approaches demonstrate that adenosine acts upon the circadian clockwork via adenosine A1/A2A receptor signalling through the activation of the Ca2+ -ERK-AP-1 and CREB/CRTC1-CRE pathways to regulate the clock genes Per1 and Per2. We show that these signalling pathways converge upon and inhibit the same pathways activated by light. Thus, circadian entrainment by light is systematically modulated on a daily basis by sleep history. These findings contribute to our understanding of how adenosine integrates signalling from both light and sleep to regulate circadian timing in mice.

Distal renal tubular acidosis: ERKNet/ESPN clinical practice points.

Distal renal tubular acidosis (dRTA) is characterized by an impaired ability of the distal tubule to excrete acid, leading to metabolic acidosis. Associated complications include bone disease, growth failure, urolithiasis and hypokalaemia. Due to its rarity, there is limited evidence to guide diagnosis and management; however, available data strongly suggest that metabolic control of the acidosis by alkali supplementation can halt or revert almost all complications. Despite this, cohort studies show that adequate metabolic control is present in only about half of patients, highlighting problems with treatment provision or adherence. With these clinical practice points the authors, part of the working groups tubulopathies in the European Rare Kidney Disease Reference network and inherited kidney diseases of the European Society for Paediatric Nephrology, aim to provide guidance for the management of patients with dRTA to facilitate adequate treatment and establish an initial best practice standard against which treatment of patients can be audited.

The circadian clock component BMAL1 regulates SARS-CoV-2 entry and replication in lung epithelial cells.

The COVID-19 pandemic, caused by SARS-CoV-2 coronavirus, is a global health issue with unprecedented challenges for public health. SARS-CoV-2 primarily infects cells of the respiratory tract, via Spike glycoprotein binding angiotensin-converting enzyme (ACE2). Circadian rhythms coordinate an organism’s response to its environment and can regulate host susceptibility to virus infection. We demonstrate a circadian regulation of ACE2 in lung epithelial cells and show that silencing BMAL1 or treatment with a synthetic REV-ERB agonist SR9009 reduces ACE2 expression and inhibits SARS-CoV-2 entry. Treating infected cells with SR9009 limits viral replication and secretion of infectious particles, showing that post-entry steps in the viral life cycle are influenced by the circadian system. Transcriptome analysis revealed that Bmal1 silencing induced a wide spectrum of interferon stimulated genes in Calu-3 lung epithelial cells, providing a mechanism for the circadian pathway to dampen SARS-CoV-2 infection. Our study suggests new approaches to understand and improve therapeutic targeting of SARS-CoV-2.

Extracorporeal treatment for calcium channel blocker poisoning: systematic review and recommendations from the EXTRIP workgroup.

BACKGROUND: Calcium channel blockers (CCBs) are commonly used to treat conditions such as arterial hypertension and supraventricular dysrhythmias. Poisoning from these drugs can lead to severe morbidity and mortality. We aimed to determine the utility of extracorporeal treatments (ECTRs) in the management of CCB poisoning. METHODS: We conducted systematic reviews of the literature, screened studies, extracted data, summarized findings, and formulated recommendations following published EXTRIP methods. RESULTS: A total of 83 publications (6 in vitro and 1 animal experiments, 55 case reports or case series, 19 pharmacokinetic studies, 1 cohort study and 1 systematic review) met inclusion criteria regarding the effect of ECTR. Toxicokinetic or pharmacokinetic data were available on 210 patients (including 32 for amlodipine, 20 for diltiazem, and 52 for verapamil). Regardless of the ECTR used, amlodipine, bepridil, diltiazem, felodipine, isradipine, mibefradil, nifedipine, nisoldipine, and verapamil were considered not dialyzable, with variable levels of evidence, while no dialyzability grading was possible for nicardipine and nitrendipine. Data were available for clinical analysis on 78 CCB poisoned patients (including 32 patients for amlodipine, 16 for diltiazem, and 23 for verapamil). Standard care (including high dose insulin euglycemic therapy) was not systematically administered. Clinical data did not suggest an improvement in outcomes with ECTR. Consequently, the EXTRIP workgroup recommends against using ECTR in addition to standard care for patients severely poisoned with either amlodipine, diltiazem or verapamil (strong recommendations, very low quality of the evidence (1D)). There were insufficient clinical data to draft recommendation for other CCBs, although the workgroup acknowledged the low dialyzability from, and lack of biological plausibility for, ECTR. CONCLUSIONS: Both dialyzability and clinical data do not support a clinical benefit from ECTRs for CCB poisoning. The EXTRIP workgroup recommends against using extracorporeal methods to enhance the elimination of amlodipine, diltiazem, and verapamil in patients with severe poisoning.

Exercise intensity-dependent effects of arm and leg-cycling on cognitive performance.

Physiological responses to arm and leg-cycling are different, which may influence psychological and biological mechanisms that influence post-exercise cognitive performance. The aim of this study was to determine the effects of maximal and submaximal (absolute and relative intensity matched) arm and leg-cycling on executive function. Thirteen males (age, 24.7 ± 5.0 years) initially undertook two incremental exercise tests to volitional exhaustion for arm-cycling (82 ± 18 W) and leg-cycling (243 ± 52 W) for the determination of maximal power output. Participants subsequently performed three 20-min constant load exercise trials: (1) arm-cycling at 50% of the ergometer-specific maximal power output (41 ± 9 W), (2) leg-cycling at 50% of the ergometer-specific maximal power output (122 ± 26 W), and (3) leg-cycling at the same absolute power output as the submaximal arm-cycling trial (41 ± 9 W). An executive function task was completed before, immediately after and 15-min after each exercise test. Exhaustive leg-cycling increased reaction time (p < 0.05, d = 1.17), while reaction time reduced following exhaustive arm-cycling (p < 0.05, d = -0.62). Improvements in reaction time were found after acute relative intensity arm (p < 0.05, d = -0.76) and leg-cycling (p < 0.05, d = -0.73), but not following leg-cycling at the same absolute intensity as arm-cycling (p > 0.05). Improvements in reaction time following arm-cycling were maintained for at least 15-min post exercise (p = 0.008, d = -0.73). Arm and leg-cycling performed at the same relative intensity elicit comparable improvements in cognitive performance. These findings suggest that individuals restricted to arm exercise possess a similar capacity to elicit an exercise-induced cognitive performance benefit.

Estimate your dose: RADDOSE-3D.

We present the current status of RADDOSE-3D, a software tool allowing the estimation of the dose absorbed in a macromolecular crystallography diffraction experiment. The code allows a temporal and spatial dose contour map to be calculated for a crystal of any geometry and size as it is rotated in an X-ray beam, and gives several summary dose values: among them diffraction weighted dose. This allows experimenters to plan data collections which will minimize radiation damage effects by spreading the absorbed dose more homogeneously, and thus to optimize the use of their crystals. It also allows quantitative comparisons between different radiation damage studies, giving a universal "x-axis" against which to plot various metrics.

Selective screening for distal renal tubular acidosis in recurrent kidney stone formers: initial experience and comparison of the simultaneous furosemide and fludrocortisone test with the short ammonium chloride test.

BACKGROUND: Distal renal tubular acidosis (dRTA) is associated with renal stone disease, and it often needs to be considered and excluded in some recurrent calcium kidney stone formers (KSFs). However, a diagnosis of dRTA, especially when 'incomplete', can be missed and needs to be confirmed by a urinary acidification (UA) test. The gold standard reference test is still the short ammonium chloride (NH4Cl) test, but it is limited by gastrointestinal side effects and occasionally failure to ingest sufficient NH4Cl. For this reason, the furosemide plus fludrocortisone (F+F) test has been proposed as an easier and better-tolerated screening test. The aim of the present study was to assess the usefulness of the F+F test as a clinical screening tool for dRTA in a renal stone clinic. METHODS: We studied 124 patients retrospectively in whom incomplete dRTA was suspected: 71 had kidney stones only, 9 had nephrocalcinosis only and 44 had both. A total of 158 UA tests were performed: 124 F+F and 34 NH4Cl; both tests were completed in 34 patients. RESULTS: The mean age was 45.4 ± 15 years, and 49% of patients were male. The prevalence of complete and incomplete dRTAs was 7 and 13.7%, respectively. Of the 34 patients tested using both tests, 17 (50%) were abnormal and 4 (12%) were normal. Thirteen (39%) patients were abnormal by F+F, but normal by NH4Cl [sensitivity 100% (95% CI 80-100), specificity 24% (95% CI 7-50), positive predictive value 57% (95% CI 37-75), negative predictive value 100% (95% CI 40-100)]. CONCLUSIONS: The F+F test is characterized by an excellent sensitivity and negative predictive value, and the diagnosis of incomplete dRTA can be excluded reliably in a patient who acidifies their urine normally with this test. However, its lack of specificity is a drawback, and if there is any doubt, an abnormal F+F test may need to be confirmed by a follow-up NH4Cl test. Ideally, a prospective blinded study in unselected KSFs is needed to accurately assess the reliability of the F+F test in diagnosing, rather than excluding, dRTA.

Outcomes in Cardiac Arrest Patients due to Toxic Exposure Treated with Therapeutic Hypothermia.

The incidence and outcome of patients who undergo therapeutic hypothermia (TH) after toxin-induced cardiac arrest (TICA) is not previously described. Our study aimed to describe the incidence, epidemiologic characteristics, and outcomes of patients who experience TICA in a dedicated clinical pathway for post-cardiac arrest care between November 2007 and February 2013. All patients were treated in an evidence-based clinical pathway that included TH. Database and medical records were independently reviewed by investigators to ascertain TICA. TICA was defined as cardiac arrest (CA) directly and immediately caused by a xenobiotic exposure. All patients were enrolled at Carolinas Medical Center, an urban 874-bed teaching hospital that serves as a regional cardiac resuscitation center. All patients were adult victims of cardiac arrest who had obtained return of spontaneous circulation and were enrolled in a clinical pathway for post-cardiac arrest care that included TH. Three hundred eighty-nine patients underwent treatment following CA during the study period and 48 (12 %) were deemed TICA. Patients who suffered TICA were slightly younger, less likely to have an initial shockable rhythm, and less likely to receive bystander CPR as compared to non-toxic cases. TICA accounted for a significant proportion of patients in this study. Additional, larger studies are needed to fully elucidate the optimal role for TH in TICA.

Cardiotoxicodynamics: Toxicity of Cardiovascular Xenobiotics.

Maintaining adequate tissue perfusion depends on a variety of factors, all of which can be influenced by xenobiotics (substances foreign to the body, including pharmaceuticals, chemicals, and natural compounds). Volume status, systemic vascular resistance, myocardial contractility, and cardiac rhythm all play a significant role in ensuring hemodynamic stability and proper cardiovascular function. Direct effects on the nervous system, the vasculature, or the heart itself as well as indirect metabolic effects may play a significant role in the development of cardiotoxicity. This article is dedicated to discussion of the disruption of cardiovascular physiology by xenobiotics.

Effect of being overweight on urinary metabolic risk factors for kidney stone formation.

BACKGROUND: The prevalence and incidence of kidney stone disease have increased markedly during the past several decades, and studies have demonstrated that inappropriate dietary habits are leading to more obesity and overweight (OW) in children and adults, which may be important in stone formation. Obese and OW patients share most of the same risk factors for cardiovascular morbidity, while the impact of being OW, rather than obese, on urinary metabolic parameters of kidney stone formers (KSF) is less well known. The aims of this study were to investigate urinary metabolic parameters, stone composition and probability of stone formation (Psf) in OW KSF when compared with normal weight (NW) and obese KSF. METHODS: The kidney stone database for KSF attending a large metabolic stone clinic was investigated. Patients with a recorded BMI, confirmed diagnosis of kidney stone disease and full metabolic evaluation were divided into three categories: BMI ≤25.0 kg/m(2) (NW group), BMI 25-30 kg/m(2) (OW group) and BMI >30.0 kg/m(2) (obese group). Twenty-four hour urinary volume (U.Vol), pH (U.pH), calcium (U.Ca), oxalate (U.Ox), citrate (U.Cit), uric acid (U.UA), magnesium (U.Mg), sodium (U.Na) and potassium (U.K) excretions, along with stone composition and Psf, were then compared among the groups. RESULTS: A total of 2132 patients were studied, of whom 833 (39%) were NW, 863 (40.5%) were OW and 436 (20.5%) were obese. OW and obese KSF were older (mean age 43 ± 15 in NW, 48 ± 13 in OW and 50 ± 12 years in obese; P for trend <0.001), demonstrated increased female predominance and higher prevalence of diabetes, hypertension and gout. There were no statistically significant differences in U.Vol and U.Mg among the groups. However, significantly higher levels of U.Ca, U.Ox, U.Cit, by crude analysis, and U.UA (3.3 ± 1.1 versus 3.8 ± 1.2 versus 4.0 ± 1.2 mmol/L; P < 0.001 for trend), U.Na (151 ± 57 versus 165 ± 60 versus 184 ± 63 mmol/L; P < 0.001 for trend), and lower U.pH (6.3 ± 0.5 versus 6.1 ± 0.5 versus 6.0 ± 0.6; P < 0.001 for trend) by both crude and multivariate adjusted analysis models were demonstrated in OW and obese KSF. Stone composition data (N = 640) showed a significantly higher incidence of uric acid stones in OW and obese groups (P for trend < 0.001). In addition, higher Psf for CaOx, UA and CaOx/UA stone types were detected in OW and obese compared with NW KSF. CONCLUSIONS: Similar to obese KSF, OW KSF show clear alterations in metabolic urinary profiles that are associated with increased overall risk of stone formation. This greater risk is primarily due to raised U.UA and U.Na, lower U.pH and higher prevalence of hypercalciuria, along with unchanged levels of the commonly measured urinary lithogenesis inhibitors. Moreover, our study established a higher incidence of uric acid, but not calcium, stones in OW KSF. Thus, appropriate evaluation and follow-up may be warranted even in OW patients who are at risk of increased stone formation. Whether modest weight loss in OW KSF will have a favourable impact on their metabolic urinary profiles and thereby diminish the risk of further stone formation needs exploring.

Intracellular light-activation of riboswitch activity.

By combining a riboswitch with a cell-permeable photocaged small-molecule ligand, an optochemical gene control element was constructed that enabled spatial and temporal control of gene expression in bacterial cells. The simplicity of this strategy, coupled with the ability to create synthetic riboswitches with tailored ligand specificities and output in a variety of microorganisms, plants, and fungi might afford a general strategy to photocontrol gene expression in vivo. The ability to activate riboswitches by using light enables the interrogation and manipulation of a wide range of biological processes with high precision, and will have broad utility in the regulation of artificial genetic circuits.

Negative predictive value of acetaminophen concentrations within four hours of ingestion.

OBJECTIVES: The objective was to ascertain whether acetaminophen (APAP) concentrations less than 100 µg/mL obtained between 1 and 4 hours after acute ingestion accurately predict a nontoxic 4-hour concentration. METHODS: The authors performed a multicenter, prospective cohort study involving five emergency departments (EDs) participating in the ToxIC Research Network. Data were collected from May 2009 to December 2011. Patients with APAP concentrations <100 µg/mL drawn between 1 and 4 hours after acute ingestions, and concentrations drawn 4 or more hours after ingestions, were included in the study. Exclusion criteria included initial concentration >100 µg/mL, initial APAP concentration drawn prior to 1 hour, two undetectable APAP concentrations, the second concentration drawn prior to 4 hours, and unknown time of ingestion. Toxic concentrations 4 or more hours after ingestion were defined as concentrations that plotted above the 150 µg/mL line on the Rumack-Matthew nomogram. RESULTS: Data were collected on 83 patients who met inclusion criteria. Of the 83 patients with APAP concentrations <100 µg/mL between 1 and 4 hours, one patient (1.2%) had a ≥ 4-hour toxic concentration. Negative predictive value (NPV) for an APAP concentration <100 µg/mL obtained between 1 and 4 hours after an acute ingestion was 98.8% (95% confidence interval [CI] = 93.5% to 99.8%). CONCLUSIONS: An APAP concentration of <100 µg/mL obtained between 1 and 4 hours after ingestion has a high NPV for excluding toxic ingestion. We do not recommend reliance on concentrations obtained between 1 and 4 hours to exclude toxicity, because of a potential false-negative rate of 6.5%.

A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications.

Polyvinyl alcohol (PVA) is a synthetic polymer derived from polyvinyl acetate through partial or full hydroxylation. PVA is commonly used in medical devices due to its low protein adsorption characteristics, biocompatibility, high water solubility, and chemical resistance. Some of the most common medical uses of PVA are in soft contact lenses, eye drops, embolization particles, tissue adhesion barriers, and as artificial cartilage and meniscus. The purpose of this review is to evaluate the available published information on PVA with respect to its safety as a medical device implant material for cartilage replacement. The review includes historical clinical use of PVA in orthopedics, and in vitro and in vivo biocompatibility studies. Finally, the safety recommendation involving the further development of PVA cryogels for cartilage replacement is addressed.

Taking it to the street: a psychiatric outreach service in Canada.

This paper describes a model of flexible psychiatric outreach service in Canada designed to meet the needs of persons who are homeless or marginally housed and have mental illness. The activities of the Psychiatric Outreach Team of the Royal Ottawa Hospital for individual clients and the community agencies who serve them are profiled, followed by a demographic and mental and physical health profile of the clients seen in the past year. The differences from other models of service and the benefits and limitations of this unique multidisciplinary team are discussed, with implications for future service development for this vulnerable population.