A Genetic Map of the Response to DNA Damage in Human Cells.

The response to DNA damage is critical for cellular homeostasis, tumor suppression, immunity, and gametogenesis. In order to provide an unbiased and global view of the DNA damage response in human cells, we undertook 31 CRISPR-Cas9 screens against 27 genotoxic agents in the retinal pigment epithelium-1 (RPE1) cell line. These screens identified 890 genes whose loss causes either sensitivity or resistance to DNA-damaging agents. Mining this dataset, we discovered that ERCC6L2 (which is mutated in a bone-marrow failure syndrome) codes for a canonical non-homologous end-joining pathway factor, that the RNA polymerase II component ELOF1 modulates the response to transcription-blocking agents, and that the cytotoxicity of the G-quadruplex ligand pyridostatin involves trapping topoisomerase II on DNA. This map of the DNA damage response provides a rich resource to study this fundamental cellular system and has implications for the development and use of genotoxic agents in cancer therapy.

A Novel Circulating MicroRNA for the Detection of Acute Myocarditis.

BACKGROUND: The diagnosis of acute myocarditis typically requires either endomyocardial biopsy (which is invasive) or cardiovascular magnetic resonance imaging (which is not universally available). Additional approaches to diagnosis are desirable. We sought to identify a novel microRNA for the diagnosis of acute myocarditis. METHODS: To identify a microRNA specific for myocarditis, we performed microRNA microarray analyses and quantitative polymerase-chain-reaction (qPCR) assays in sorted CD4+ T cells and type 17 helper T (Th17) cells after inducing experimental autoimmune myocarditis or myocardial infarction in mice. We also performed qPCR in samples from coxsackievirus-induced myocarditis in mice. We then identified the human homologue for this microRNA and compared its expression in plasma obtained from patients with acute myocarditis with the expression in various controls. RESULTS: We confirmed that Th17 cells, which are characterized by the production of interleukin-17, are a characteristic feature of myocardial injury in the acute phase of myocarditis. The microRNA mmu-miR-721 was synthesized by Th17 cells and was present in the plasma of mice with acute autoimmune or viral myocarditis but not in those with acute myocardial infarction. The human homologue, designated hsa-miR-Chr8:96, was identified in four independent cohorts of patients with myocarditis. The area under the receiver-operating-characteristic curve for this novel microRNA for distinguishing patients with acute myocarditis from those with myocardial infarction was 0.927 (95% confidence interval, 0.879 to 0.975). The microRNA retained its diagnostic value in models after adjustment for age, sex, ejection fraction, and serum troponin level. CONCLUSIONS: After identifying a novel microRNA in mice and humans with myocarditis, we found that the human homologue (hsa-miR-Chr8:96) could be used to distinguish patients with myocarditis from those with myocardial infarction. (Funded by the Spanish Ministry of Science and Innovation and others.).

Scalable protein production by Komagataella phaffii enabled by ARS plasmids and carbon source-based selection.

BACKGROUND: Most recombinant Komagataella phaffii (Pichia pastoris) strains for protein production are generated by genomic integration of expression cassettes. The clonal variability in gene copy numbers, integration loci and consequently product titers limit the aptitude for high throughput applications in drug discovery, enzyme engineering or most comparative analyses of genetic elements such as promoters or secretion signals. Circular episomal plasmids with an autonomously replicating sequence (ARS), an alternative which would alleviate some of these limitations, are inherently unstable in K. phaffii. Permanent selection pressure, mostly enabled by antibiotic resistance or auxotrophy markers, is crucial for plasmid maintenance and hardly scalable for production. The establishment and use of extrachromosomal ARS plasmids with key genes of the glycerol metabolism (glycerol kinase 1, GUT1, and triosephosphate isomerase 1, TPI1) as selection markers was investigated to obtain a system with high transformation rates that can be directly used for scalable production processes in lab scale bioreactors. RESULTS: In micro-scale deep-well plate experiments, ARS plasmids employing the Ashbya gossypii TEF1 (transcription elongation factor 1) promoter to regulate transcription of the marker gene were found to deliver high transformation efficiencies and the best performances with the reporter protein (CalB, lipase B of Candida antarctica) for both, the GUT1- and TPI1-based, marker systems. The GUT1 marker-bearing strain surpassed the reference strain with integrated expression cassette by 46% upon re-evaluation in shake flask cultures regarding CalB production, while the TPI1 system was slightly less productive compared to the control. In 5 L bioreactor methanol-free fed-batch cultivations, the episomal production system employing the GUT1 marker led to 100% increased CalB activity in the culture supernatant compared to integration construct. CONCLUSIONS: For the first time, a scalable and methanol-independent expression system for recombinant protein production for K. phaffii using episomal expression vectors was demonstrated. Expression of the GUT1 selection marker gene of the new ARS plasmids was refined by employing the TEF1 promoter of A. gossypii. Additionally, the antibiotic-free marker toolbox for K. phaffii was expanded by the TPI1 marker system, which proved to be similarly suited for the use in episomal plasmids as well as integrative expression constructs for the purpose of recombinant protein production.

Pregnancy, Reproductive Factors, and Female Heart Failure Risk and Outcomes.

PURPOSE OF REVIEW: The purpose of this review is to provide an overview of recent evidence on female-specific risk factors related to reproductive status or pregnancy. RECENT FINDINGS: Pregnancy-related factors, including hypertensive disorders and gestational diabetes, increase the risk of heart failure in women, while breastfeeding and hormone therapy may offer protection. Hypertensive disorders of pregnancy, gestational diabetes, polycystic ovarian syndrome, placental abruption, younger maternal age at first live birth, younger maternal age at last live birth, number of stillbirths, number of pregnancies, onset of menstruation before 12 years of age, shorter reproductive age, ovariectomy, and prolonged absence of ovarian hormones may increase the risk of heart failure in women. Conversely, breastfeeding status and hormone therapy (for menopause or contraception) may serve as protective factors, while fertility treatments have no discernible effect on the risk of heart failure.

Effects of intravenous lysine acetylsalicylate versus oral aspirin on platelet responsiveness in patients with ST-segment elevation myocardial infarction: the ECCLIPSE-STEMI trial.

Prasugrel and ticagrelor, new P2Y12-ADP receptor antagonists, are associated with greater pharmacodynamic inhibition and reduction of cardiovascular events in patients with an acute coronary syndrome. However, evidence is lacked about the effects of achieving faster and stronger cyclooxygenase inhibition with intravenous lysine acetylsalicylate (LA) compared to oral aspirin. Recently, we demonstrated in healthy volunteers that the administration of intravenous LA resulted in a significantly reduction of platelet reactivity compared to oral aspirin. Loading dose of LA achieves platelet inhibition faster, and with less variability than aspirin. However, there are no data of this issue in patients with an ST-segment elevation myocardial infarction (STEMI). This is a prospective, randomized, multicenter, open platelet function study conducted in STEMI patients. Subjects were randomly assigned to receive a loading dose (LD) of intravenous LA 450 mg plus oral ticagrelor 180 mg, or LD of aspirin 300 mg plus ticagrelor 180 mg orally. Platelet function was evaluated at baseline, 30 min, 1 h, 4 h and 24 h using multiple electrode aggregometry and vasodilator-stimulated phosphoprotein phosphorylation (VASP). The primary endpoint of the study is the inhibition of platelet aggregation (IPA) after arachidonic acid (AA) 0.5 mM at 30 min. Secondary endpoints were the IPA at 1, 4, and 24 h after AA, and non-AA pathways through the sequence (ADP and TRAP). A total of 32 STEMI patients were randomized (16 LA, 16 aspirin). The inhibition of platelet aggregation after AA 0.5 mM at 30 min was greater in subjects treated with LA compared with aspirin: 166 vs. 412 respectively (p = 0.001). This differential effect was observed at 1 h (p = 0.01), but not at 4 and 24 h. Subjects treated with LA presented less variability and faster inhibition of platelet aggregation wit AA compared with aspirin. The administration of intravenous LA resulted in a significantly reduction of platelet reactivity compared to oral aspirin on ticagrelor inhibited platelets in patients with STEMI. Loading dose of LA achieves an earlier platelet inhibition, and with less variability than aspirin.Trial Registration: Unique identifier: NCT02929888; URL: http://www.clinicaltrials.gov.

Serially improved GTOs for molecular applications (SIGMA): Basis sets from H to Ne.

A new approach for generating Gaussian basis sets is reported and tested for atoms from H to Ne. The basis sets thus calculated, named SIGMA basis sets, range from DZ to QZ sizes and have the same composition per shell as Dunning basis sets but with different treatment of the contractions. The standard SIGMA basis sets and their augmented versions have proven to be very suitable for providing good results in atomic and molecular calculations. The performance of the new basis sets is analyzed in terms of total, correlation, and atomization energies, equilibrium distances, and vibrational frequencies in several molecules, and the results are compared at several computational levels with those obtained with the corresponding Dunning and other basis sets.

Emotional empowerment through information research and communication to reduce fear of COVID-19 among nursing students: a randomised controlled trial.

BACKGROUND: Despite an existing body of literature on anxiety reduction using multi-component methods, little is known about the effect of active student participation in research and communication of scientific information on anxiety and fear reduction. The aim of this study is to evaluate the impact of quality scientific information research and the production of informative videos on the preventive aspects of COVID-19 on fear and anxiety reduction. METHODS: A randomised controlled trial was conducted with 220 undergraduate nursing students in the first year of the nursing degree. The participating students were randomised into two groups. The experimental group carried out an intervention based on a database search for information on preventing COVID-19 and production of a video giving scientific reasons why prevention measures should be followed. In the control group, students created posters and videos about theoretical aspects of one module of the nursing degree. Both groups were surveyed before and after the intervention, measuring their state of resilience, preventive behaviours, level of anxiety, and fear of COVID-19. RESULTS: The intervention group showed a greater decrease in fear levels after the intervention than those in the control group. There were no differences between the groups in terms of resilience, preventive behaviours, or anxiety. In the experimental group, there was a significant decrease in anxiety levels and fear levels after the intervention compared to the baseline levels. CONCLUSIONS: An intervention based on active participation in searches for high-quality scientific information and production of information videos on COVID-19 prevention reduced fear and anxiety caused by COVID-19 among nursing students. TRIAL REGISTRATION: We have retrospectively registered the trial in Open Science Framework and the identification number is https://doi.org/10.17605/OSF.IO/6QU5S .

Efficacy-safety profile of the video capsule endoscopy in the study of the small bowel: experience over 100 consecutive procedures.

Video capsule endoscopy (VCE) is currently considered the first-line study in the evaluation of the small bowel (SB). Retrospective study including consecutive patients from 2010 to 2021 in two referral endoscopic centers in Peru, who underwent VCE. Inclusion criteria were patients with middle gastrointestinal bleeding, chronic diarrhea and unexplained chronic abdominal pain (endoscopic studies prior to VCE: normal). We mainly used Pillcam SB VCE (Given Imaging, Israel) SB2 and SB3 generations.

Sensing Enhancement on Social Networks: The Role of Network Topology.

Sensing and processing information from dynamically changing environments is essential for the survival of animal collectives and the functioning of human society. In this context, previous work has shown that communication between networked agents with some preference towards adopting the majority opinion can enhance the quality of error-prone individual sensing from dynamic environments. In this paper, we compare the potential of different types of complex networks for such sensing enhancement. Numerical simulations on complex networks are complemented by a mean-field approach for limited connectivity that captures essential trends in dependencies. Our results show that, whilst bestowing advantages on a small group of agents, degree heterogeneity tends to impede overall sensing enhancement. In contrast, clustering and spatial structure play a more nuanced role depending on overall connectivity. We find that ring graphs exhibit superior enhancement for large connectivity and that random graphs outperform for small connectivity. Further exploring the role of clustering and path lengths in small-world models, we find that sensing enhancement tends to be boosted in the small-world regime.

Diagnosis, prevention, and management of delirium in the intensive cardiac care unit.

Delirium is a frequent complication in patients admitted to intensive cardiac care units (ICCU) with potentially severe consequences including increased risks of mortality, cognitive impairment and dependence at discharge, and longer times on mechanical ventilation and hospital stay. Delirium has been widely documented and studied in general intensive care units and in patients after cardiac surgery, but it has barely been studied in acute nonsurgical cardiac patients. Moreover, delirium (especially in its hypoactive form) is commonly misdiagnosed. We propose a protocol for delirium prevention and management in ICCUs. A daily comprehensive assessment to improve detection should be done using validated scales (ie, confusion assessment method). Preventive measures are particularly relevance and constitute the basis of treatment as well, acting on reversible risk factors, including environmental interventions, such as quiet time, sleep promotion, family support, communication, and adequate treatment of pain and dyspnea. Pharmacological prophylaxis is not indicated with the exception of patients at risk of withdrawal syndrome but should only be used in patients with confirmed delirium. Dexmedetomidine is the drug of choice in patients with severe agitation, and those weaning from invasive mechanical ventilation. As the complexity of ICCUs increases, clinical scenarios posing challenges for the management of delirium become more frequent. Efforts should be done to improve the identification of patients at risk during admission in order to establish preventive interventions to avoid this complication. Patient-centered protocols will increase the awareness of the healthcare professionals for better prevention and earlier diagnosis and will positively impact on prognosis.

Intra-Day Solar Power Forecasting Strategy for Managing Virtual Power Plants.

Solar energy penetration has been on the rise worldwide during the past decade, attracting a growing interest in solar power forecasting over short time horizons. The increasing integration of these resources without accurate power forecasts hinders the grid operation and discourages the use of this renewable resource. To overcome this problem, Virtual Power Plants (VPPs) provide a solution to centralize the management of several installations to minimize the forecasting error. This paper introduces a method to efficiently produce intra-day accurate Photovoltaic (PV) power forecasts at different locations, by using free and available information. Prediction intervals, which are based on the Mean Absolute Error (MAE), account for the forecast uncertainty which provides additional information about the VPP node power generation. The performance of the forecasting strategy has been verified against the power generated by a real PV installation, and a set of ground-based meteorological stations in geographical proximity have been used to emulate a VPP. The forecasting approach is based on a Long Short-Term Memory (LSTM) network and shows similar errors to those obtained with other deep learning methods published in the literature, offering a MAE performance of 44.19 W/m2 under different lead times and launch times. By applying this technique to 8 VPP nodes, the global error is reduced by 12.37% in terms of the MAE, showing huge potential in this environment.

[Clinical practice guidelines for diagnosis and management of Helicobacter pylori infection in gastroduodenal diseases of the Peruvian Health Social Security (EsSalud)]. / Guía de práctica clínica para el diagnóstico y manejo de la infección por Helicobacter pylori en enfermedades gastroduodenales en el Seguro Social de Salud del Perú (EsSalud).

INTRODUCTION: This article summarizes the evidence-based recommendations of the clinical practice guide (CPG) for the diagnosis and management of Helicobacter pylori infection in gastroduodenal diseases. METHODS: For the provision of these recommendations, a guideline development group (local GDG) was established, including medical specialists andmethodologists that formulated seven clinical questions. Systematic searches of systematic reviews and -when it was considered pertinent- primary studies were conducted in PubMed and CENTRAL during December 2017 and July 2019. The evidence to answer each of the posed clinical questions was selected. The quality of the evidence was evaluated using the Grading ofRecommendations Assessment, Development, and Evaluation (GRADE) methodology. In periodic work meetings, the local GDG used the GRADE methodology to review the evidence and formulate the recommendations, points of good clinical practice, and flowcharts. Finally, the CPG was approved with Resolution N° 104-IETSI-ESSALUD-2020. RESULTS: This CPG addressed seven clinical questions, divided into four topics. Based on these questions, 12 recommendations (3 strong and 9 weak), 17 points of good clinical practice, and two flowcharts (one for diagnosis and another for management) were formulated. CONCLUSION: This article summarizes the methodology and evidence-based conclusions from the CPG for for the diagnosis and management of Helicobacter pylori infection in gastroduodenal diseases.

Polµ tumor variants decrease the efficiency and accuracy of NHEJ.

The non homologous end-joining (NHEJ) pathway of double-strand break (DSB) repair often requires DNA synthesis to fill the gaps generated upon alignment of the broken ends, a complex task performed in human cells by two specialized DNA polymerases, Polλ and Polµ. It is now well established that Polµ is the one adapted to repair DSBs with non-complementary ends, the most challenging scenario, although the structural basis and physiological implications of this adaptation are not fully understood. Here, we demonstrate that two human Polµ point mutations, G174S and R175H, previously identified in two different tumor samples and affecting two adjacent residues, limit the efficiency of accurate NHEJ by Polµ in vitro and in vivo. Moreover, we show that this limitation is the consequence of a decreased template dependency during NHEJ, which renders the error-rate of the mutants higher due to the ability of Polµ to randomly incorporate nucleotides at DSBs. These results highlight the relevance of the 8 kDa domain of Polµ for accurate and efficient NHEJ, but also its contribution to the error-prone behavior of Polµ at 2-nt gaps. This work provides the first demonstration that mutations affecting Polµ identified in tumors can alter the efficiency and fidelity of NHEJ.

A Hybrid Approach to Short-Term Load Forecasting Aimed at Bad Data Detection in Secondary Substation Monitoring Equipment.

Bad data as a result of measurement errors in secondary substation (SS) monitoring equipment is difficult to detect and negatively affects power system state estimation performance by both increasing the computational burden and jeopardizing the state estimation accuracy. In this paper a short-term load forecasting (STLF) hybrid strategy based on singular spectrum analysis (SSA) in combination with artificial neural networks (ANN), is presented. This STLF approach is aimed at detecting, identifying and eliminating and/or correcting such bad data before it is provided to the state estimator. This approach is developed to improve the accuracy of the load forecasts and it is tested against real power load data provided by electricity suppliers. Depending on the week considered, mean absolute percentage error (MAPE) values which range from 1.6% to 3.4% are achieved for STLF. Different systematic errors, such as gain and offset error levels and outliers, are successfully detected with a hit rate of 98%, and the corresponding measurements are corrected before they are sent to the control center for state estimation purposes.

Management of late presentation congenital heart disease.

In many parts of the world, mostly low- and middle-income countries, timely diagnosis and repair of congenital heart diseases (CHDs) is not feasible for a variety of reasons. In these regions, economic growth has enabled the development of cardiac units that manage patients with CHD presenting later than would be ideal, often after the window for early stabilisation - transposition of the great arteries, coarctation of the aorta - or for lower-risk surgery in infancy - left-to-right shunts or cyanotic conditions. As a result, patients may have suffered organ dysfunction, manifest signs of pulmonary vascular disease, or the sequelae of profound cyanosis and polycythaemia. Late presentation poses unique clinical and ethical challenges in decision making regarding operability or surgical candidacy, surgical strategy, and perioperative intensive care management.

ATP insertion opposite 8-oxo-deoxyguanosine by Pol4 mediates error-free tolerance in Schizosaccharomyces pombe.

7,8-Dihydro-8-oxo-deoxyguanosine (8oxodG) is a highly premutagenic DNA lesion due to its ability to mispair with adenine. Schizosaccharomyces pombe lacks homologs for relevant enzymes that repair 8oxodG, which suggests that this lesion could be persistent and must be tolerated. Here we show that SpPol4, the unique PolX in fission yeast, incorporates ATP opposite 8oxodG almost exclusively when all nucleotides (ribos and deoxys) are provided at physiological concentrations. Remarkably, this SpPol4-specific reaction could also occur during the NHEJ of DSBs. In cell extracts, misincorporation of ATP opposite 8oxodG was shown to be SpPol4-specific, although RNase H2 efficiently recognized the 8oxodG:AMP mispair to remove AMP and trigger error-free incorporation of dCTP. These data are the first evidence that ribonucleotides can be used safely for 8oxodG tolerance, suggesting that insertion of the highly abundant ATP substrate could be beneficial to promote efficient and error-free repair of 8oxodG-associated DSBs. Moreover, we demonstrate that purified SpPol4 uses 8oxo-dGTP and 8oxo-GTP as substrates for DNA polymerization, although with poor efficiency compared to the incorporation of undamaged nucleotides opposite either 8oxodG or undamaged templates. This suggests that SpPol4 is specialized in tolerating 8oxodG as a DNA template, without contributing significantly to the accumulation of this lesion in the DNA.

Yeast pol4 promotes tel1-regulated chromosomal translocations.

DNA double-strand breaks (DSBs) are one of the most dangerous DNA lesions, since their erroneous repair by nonhomologous end-joining (NHEJ) can generate harmful chromosomal rearrangements. PolX DNA polymerases are well suited to extend DSB ends that cannot be directly ligated due to their particular ability to bind to and insert nucleotides at the imperfect template-primer structures formed during NHEJ. Herein, we have devised genetic assays in yeast to induce simultaneous DSBs in different chromosomes in vivo. The repair of these breaks in trans could result in reciprocal chromosomal translocations that were dependent on classical Ku-dependent NHEJ. End-joining events leading to translocations were mainly based on the formation of short base pairing between 3'-overhanging DNA ends coupled to gap-filling DNA synthesis. A major proportion of these events were specifically dependent on yeast DNA polymerase Pol4 activity. In addition, we have discovered that Pol4-Thr(540) amino acid residue can be phosphorylated by Tel1/ATM kinase, which could modulate Pol4 activity during NHEJ. Our data suggest that the role of Tel1 in preventing break-induced chromosomal translocations can, to some extent, be due to its stimulating effect on gap-filling activity of Pol4 to repair DSBs in cis. Overall, this work provides further insight to the molecular mechanisms of DSB repair by NHEJ and presents a new perspective to the understanding of how chromosomal translocations are formed in eukaryotic cells.

[Epidemiological and clinical characteristics of inflammatory bowel disease in a tertiary referral hospital in Lima-Peru]. / Características epidemiológicas y clínicas de la enfermedad inflamatoria intestinal en un hospital de referencia de Lima-Perú.

OBJECTIVE: To identify sociodemographic, clinical, and endoscopic characteristics in patients with inflammatory bowel disease (IBD). MATERIALS AND METHODS: The study period was from January 2004 to December 2014. The final diagnosis was determined by clinical gastroenterologists experienced in the diagnosis and management of IBD, based on internationally accepted diagnostic criteria. RESULTS: 105 patients with IBD were studied, 77% with ulcerative colitis (UC) and 23% with Crohn's disease (CD). The average age of initial diagnosis for UC and CD was 53.02 and 57.7 years, respectively. Regarding the gender distribution, the male:female ratio was 1.3:1 for UC and 2:1 for CD. Predominant clinical manifestations were: diarrhea (76.5%) in CU and lower gastrointestinal bleeding / abdominal pain (66.6% for each symptom) in EC. The predominant form of presentation was moderate for both CU (49.3%) and EC (62.5%). 47% of patients with UC had extensive colitis and 54.2% of patients with CD had Ileocolitis. 6.2% of the UC patients underwent surgery, whereas 50% of the CD patients required it. CONCLUSIONS: There is a tendency to an increased detection of cases of Crohn's disease in our country and in Latin America with respect to previous studies. There is a prevalence of moderate forms of presentation for both UC and CD, and high percentages of surgery in EC is evident.

Tumor necrosis factor-α inhibition attenuates middle cerebral artery remodeling but increases cerebral ischemic damage in hypertensive rats.

Hypertension causes vascular inflammation evidenced by an increase in perivascular macrophages and proinflammatory cytokines in the arterial wall. Perivascular macrophage depletion reduced tumor necrosis factor (TNF)-α expression in cerebral arteries of hypertensive rats and attenuated inward remodeling, suggesting that TNF-α might play a role in the remodeling process. We hypothesized that TNF-α inhibition would improve middle cerebral artery (MCA) structure and reduce damage after cerebral ischemia in hypertensive rats. Six-week-old male stroke-prone spontaneously hypertensive rats (SHRSP) were treated with the TNF-α inhibitor etanercept (ETN; 1.25 mg·kg(-1)·day(-1) ip daily) or PBS (equivolume) for 6 wk. The myogenic tone generation, postischemic dilation, and passive structure of MCAs were assessed by pressure myography. Cerebral ischemia was induced by MCA occlusion (MCAO). Myogenic tone was unchanged, but MCAs from SHRSP + ETN had larger passive lumen diameter and reduced wall thickness and wall-to-lumen ratio. Cerebral infarct size was increased in SHRSP + ETN after transient MCAO, despite an improvement in dilation of nonischemic MCA. The increase in infarct size was linked to a reduction in the number of microglia in the infarct core and upregulation of markers of classical macrophage/microglia polarization. There was no difference in infarct size after permanent MCAO or when untreated SHRSP subjected to transient MCAO were given ETN at reperfusion. Our data suggests that TNF-α inhibition attenuates hypertensive MCA remodeling but exacerbates cerebral damage following ischemia/reperfusion injury likely due to inhibition of the innate immune response of the brain.