Search

The first search for the lepton number violating decay K+→π0π−μ+e+ and lepton flavour violating decays K+→π0π+μ−e+, K+→π0π+μ+e− has been performed using a dataset collected by the NA62 experiment at CERN in 2016–2018. Upper limits of 2.9×10−10, 3.1×10−10 and 5.0×10−10, respectively, are obtained at 90% CL for the branching ratios of the three decays on the assumption of uniform phase-space distributions.

Biallelic variants in the SPG11 gene account for the most common form of autosomal recessive hereditary spastic paraplegia characterized by motor and cognitive impairment, with currently no therapeutic option. We previously observed in a Spg11 knockout mouse that neurodegeneration is associated with accumulation of gangliosides in lysosomes. To test whether a substrate reduction therapy could be a therapeutic option, we downregulated the key enzyme involved in ganglioside biosynthesis using an AAV-PHP.eB viral vector expressing a miRNA targeting St3gal5. Downregulation of St3gal5 in Spg11 knockout mice prevented the accumulation of gangliosides, delayed the onset of motor and cognitive symptoms, and prevented the upregulation of serum levels of neurofilament light chain, a biomarker widely used in neurodegenerative diseases. Importantly, similar results were observed when Spg11 knockout mice were administrated venglustat, a pharmacological inhibitor of glucosylceramide synthase expected to decrease ganglioside synthesis. Downregulation of St3gal5 or venglustat administration in Spg11 knockout mice strongly decreased the formation of axonal spheroids, previously associated with impaired trafficking. Venglustat had similar effect on cultured human SPG11 neurons. In conclusion, this work identifies the first disease-modifying therapeutic strategy in SPG11, and provides data supporting its relevance for therapeutic testing in SPG11 patients.

CRISPR-Cas9-mediated gene editing has vast applications in basic and clinical research and is a promising tool for several disorders. Our lab previously developed a non-integrating RNA virus, measles virus (MeV), as a single-cycle reprogramming vector by replacing the viral attachment protein with the reprogramming factors for induced pluripotent stem cell generation. Encouraged by the MeV reprogramming vector efficiency, in this study, we develop a single-cycle MeV vector to deliver the gRNA(s) and Cas9 nuclease to human cells for efficient gene editing. We show that the MeV vector achieved on-target gene editing of the reporter (mCherry) and endogenous genes (HBB and FANCD1) in human cells. Additionally, the MeV vector achieved precise knock-in via homology-directed repair using a single-stranded oligonucleotide donor. The MeV vector is a new and flexible platform for gene knock-out and knock-in modifications in human cells, capable of incorporating new technologies as they are developed.

Summary: The human transcriptome predominantly consists of noncoding RNAs (ncRNAs), transcripts that do not encode proteins. The noncoding transcriptome governs a multitude of pathophysiological processes, offering a rich source of next-generation biomarkers. Toward achieving a holistic view of disease, the integration of these transcripts with clinical records and additional data from omic technologies (“multiomic” strategies) has motivated the adoption of artificial intelligence (AI) approaches. Given their intricate biological complexity, machine learning (ML) techniques are becoming a key component of ncRNA-based research. This article presents an overview of the potential and challenges associated with employing AI/ML-driven approaches to identify clinically relevant ncRNA biomarkers and to decipher ncRNA-associated pathogenetic mechanisms. Methodological and conceptual constraints are discussed, along with an exploration of ethical considerations inherent to AI applications for healthcare and research. The ultimate goal is to provide a comprehensive examination of the multifaceted landscape of this innovative field and its clinical implications.

A sample of 3984 candidates of the K+→π+γγ decay, with an estimated background of 291±14 events, was collected by the NA62 experiment at CERN during 2017–2018. In order to describe the observed di-photon mass spectrum, the next-to-leading order contribution in chiral perturbation theory was found to be necessary. The decay branching ratio in the full kinematic range is measured to be (9.61±0.17)×10−7. The first search for production and prompt decay of an axion-like particle with gluon coupling in the process K+→π+a, a→γγ is also reported.

RNA editing, a common and potentially highly functional form of RNA modification, encompasses two different RNA modifications, namely adenosine to inosine (A-to-I) and cytidine to uridine (C-to-U) editing. As inosines are interpreted as guanosines by the cellular machinery, both A-to-I and C-to-U editing change the nucleotide sequence of the RNA. Editing events in coding sequences have the potential to change the amino acid sequence of proteins, whereas editing events in noncoding RNAs can, for example, affect microRNA target binding. With advancing RNA sequencing technology, more RNA editing events are being discovered, studied, and reported. However, RNA editing events are still often overlooked or discarded as sequence read quality defects. With this position paper, we aim to provide guidelines and recommendations for the detection, validation, and follow-up experiments to study RNA editing, taking examples from the fields of cardiovascular and brain disease. We discuss all steps, from sample collection, storage, and preparation, to different strategies for RNA sequencing and editing-sensitive data analysis strategies, to validation and follow-up experiments, as well as potential pitfalls and gaps in the available technologies. This paper may be used as an experimental guideline for RNA editing studies in any disease context.

Atherosclerotic disease is a major cause of acute cardiovascular events. A deeper understanding of its underlying mechanisms will allow advancing personalized and patient-centered healthcare. Transcriptomic research has proven to be a powerful tool for unravelling the complex molecular pathways that drive atherosclerosis. However, low reproducibility of research findings and lack of standardization of procedures pose significant challenges in this field. In this review, we discuss how transcriptomic research can help in understanding the different phenotypes of the atherosclerotic plaque that contribute to the development and progression of atherosclerosis. We highlight the methodological challenges that need to be addressed to improve research outputs, and emphasize the importance of research protocols harmonization. We also discuss recent advances in transcriptomic research, including bulk or single-cell sequencing, and their added value in plaque phenotyping. Finally, we explore how integrated multiomics data and machine learning improve understanding of atherosclerosis and provide directions for future research.

The first search for ultra-rare K+ decays into the π+e+e−e+e− final state is reported, using a dataset collected by the NA62 experiment at CERN in 2017–2018. An upper limit of 1.4×10−8 at 90% CL is obtained for the branching ratio of the K+→π+e+e−e+e− decay, predicted in the Standard Model to be (7.2±0.7)×10−11. Upper limits at 90% CL are obtained at the level of 10−9 for the branching ratios of two prompt decay chains involving pair-production of hidden-sector mediators: K+→π+aa, a→e+e− and K+→π+S, S→A′A′, A′→e+e−.

Summary: Background: Circadian (24-h) rhythms are important regulators in physiology and disease, but systemic disease may disrupt circadian rhythmicity. Heart failure (HF) is a systemic disease affecting hormonal regulation. We investigate whether HF affects the rhythmic expression of melatonin and cortisol, main endocrine products of the central clock, and cardiac-specific troponin in patients. We corroborate the functionality of the peripheral clock directly in the organs of translational models, inaccessible in human participants. Methods: We included 46 HF patients (71.7% male, median age of 60 years, NYHA class II (32.6%) or III (67.4%), ischemic cardiomyopathy (43.5%), comorbidities: diabetes 21.7%, atrial fibrillation 30.4%), and 24 matched controls. Blood was collected at seven time-points during a 24-h period (totalling 320 HF and 167 control samples) for melatonin, cortisol, and cardiac troponin T (cTnT) measurements after which circadian rhythms were assessed through cosinor analyses, both on the individual and the group level. Next, we analysed peripheral circadian clock functionality using cosinor analysis in male animal HF models: nocturnal mice and diurnal zebrafish, based on expression of core clock genes in heart, kidneys, and liver, every 4 h during a 24-h period in a light/darkness synchronised environment. Findings: Melatonin and cortisol concentrations followed a physiological 24-h pattern in both patients and controls. For melatonin, acrophase occurred during the night for both groups, with significantly decreased amplitude (median 5.2 vs 8.8, P = 0.0001) and circadian variation ([maximum]/[minimum]) in heart failure patients. For cortisol, mesor showed a significant increase for HF patients (mean 331.9 vs 275.1, P = 0.017) with a difference of 56.8 (95% CI 10.3–103.3) again resulting in a relatively lower variation: median 3.9 vs 6.3 (P = 0.0058). A nocturnal blood pressure dip was absent in 77.8% of HF patients.Clock gene expression profiles (Bmal, Clock, Per, Cry) were similar and with expected phase relations in animal HF models and controls, demonstrating preserved peripheral clock functionality in HF. Furthermore, oscillations in diurnal zebrafish were expectedly in opposite phases to those of nocturnal mice. Concordantly, cTnT concentrations in HF patients revealed significant circadian oscillations. Interpretation: Central clock output is dampened in HF patients while the molecular peripheral clock, as confirmed in animal models, remains intact. This emphasises the importance of taking timing into account in research and therapy for HF, setting the stage for another dimension of diagnostic, prognostic and therapeutic approaches. Funding: Hartstichting.

OCT4 is a key mediator of induced pluripotent stem cell (iPSC) reprogramming, but the mechanistic insights into the role of exogenous OCT4 and timelines that initiate pluripotency remain to be resolved. Here, using measles reprogramming vectors, we present microRNA (miRNA) targeting of exogenous OCT4 to shut down its expression during the mesenchymal to the epithelial transition phase of reprogramming. We showed that exogenous OCT4 is required only for the initiation of reprogramming and is dispensable for the maturation stage. However, the continuous expression of SOX2, KLF4, and c-MYC is necessary for the maturation stage of the iPSC. Additionally, we demonstrate a novel application of miRNA targeting in a viral vector to contextually control the vector/transgene, ultimately leading to an improved reprogramming efficiency. This novel approach could be applied to other systems for improving the efficiency of vector-induced processes.

Despite promising findings, quantitative PCR (qPCR)-based tests for RNA quantification have experienced serious limitations in their clinical application. The noticeable lack of technical standardization remains a huge obstacle in the translation of qPCR-based tests. The incorporation of qPCR-based tests into the clinic will benefit from guidelines for clinical research assay validation. This will ultimately impact the clinical management of the patient, including diagnosis, prognosis, prediction, monitoring of the therapeutic response, and evaluation of toxicity. However, clear assay validation protocols for biomarker investigation in clinical trials using molecular assays are currently lacking. Here, we will focus on the necessary steps, including sample acquisition, processing and storage, RNA purification, target selection, assay design, and experimental design, that need to be taken toward the appropriate validation of qRT-PCR assays in clinical research. These recommendations can fill the gap between research use only (RUO) and in vitro diagnostics (IVD). Our contribution provides a tool for basic and clinical research for the development of validated assays in the intermediate steps of biomarker research. These guidelines are based on the current understanding and consensus within the EU-CardioRNA COST Action consortium (www.cardiorna.eu). Their applicability encompasses all clinical areas.

A search for the K+→μ−νe+e+ decay, forbidden within the Standard Model by either lepton number or lepton flavour conservation depending on the flavour of the emitted neutrino, has been performed using the dataset collected by the NA62 experiment at CERN in 2016–2018. An upper limit of 8.1×10−11 is obtained for the decay branching fraction at 90% CL, improving by a factor of 250 over the previous search.

Summary: Cardiovascular disorders such as heart failure are leading causes of mortality. Patient stratification via identification of novel biomarkers could improve management of cardiovascular diseases of complex etiologies. Long-noncoding RNAs (lncRNAs) are highly tissue-specific in nature and have emerged as important biomarkers in human diseases. In this study, we aimed to identify cardiac-enriched lncRNAs as potential biomarkers for cardiovascular conditions.Deep RNA sequencing and quantitative PCR identified differentially expressed lncRNAs between failing and non-failing hearts. An independent dataset was used to evaluate the enrichment of lncRNAs in normal hearts.We identified a panel of 2906 lncRNAs, named FIMICS, that were either cardiac-enriched or differentially expressed between failing and non-failing hearts. Expression of lncRNAs in blood samples differentiated patients with myocarditis and acute myocardial infarction.We hereby present the FIMICS panel, a readily available tool to provide insights into cardiovascular pathologies and which could be helpful for diagnosis, monitoring and prognosis purposes.

SARS-CoV-2, the virus responsible for COVID-19 in humans, can efficiently infect a large number of animal species. Like any virus, and particularly RNA viruses, SARS-CoV-2 undergoes mutations during its life cycle some of which bring a selective advantage, leading to the selection of a given lineage. Minks are very susceptible to SARS-CoV-2 and owing to their presence in mass rearing, they make a good model for studying the relative importance of mutations in viral adaptation to host species. Variants, such as the mink-selected SARS-CoV-2 Y453F and D614G or H69del/V70del, Y453F, I692V and M1229I were identified in humans after spreading through densely caged minks. However, not all mink-specific mutations are conserved when the virus infects human populations back. Many questions remain regarding the interspecies evolution of SARS-CoV-2 and the dynamics of transmission leading to the emergence of new variant strains. We compared the human and mink ACE2 receptor structures and their interactions with SARS-CVoV-2 variants. In minks, ACE2 presents a Y34 amino acid instead of the H34 amino acid found in the human ACE2. H34 is essential for the interaction with the Y453 residue of the SARS-CoV-2 Spike protein. The Y453F mink mutation abolishes this conflict. A series of 18 mutations not involved in the direct ACE2 interaction was observed in addition to the Y453F and D614G in 16 different SARS-CoV-2 strains following bidirectional infections between humans and minks. These mutations were not random and were distributed into five different functional groups having an effect on the kinetics of ACE2-RD interaction. The interspecies transmission of SARS-CoV-2 from humans to minks and back to humans, generated specific mutations in each species which improved the affinity for the ACE2 receptor either by direct mutation of the core 453 residue or by associated compensatory mutations.

Although systemic inflammation and pulmonary complications increase the mortality rate in COVID-19, a broad spectrum of cardiovascular and neurological complications can also contribute to significant morbidity and mortality.The molecular mechanisms underlying cardiovascular and neurological complications during and after SARS-CoV-2 infection are incompletely understood. Recently reported perturbations of the epitranscriptome of COVID-19 patients indicate that mechanisms including those derived from RNA modifications and non-coding RNAs may play a contributing role in the pathogenesis of COVID-19.In this review paper, we gathered recently published studies investigating (epi)transcriptomic fluctuations upon SARS-CoV-2 infection, focusing on the brain-heart axis since neurological and cardiovascular events and their sequelae are of utmost prevalence and importance in this disease.

Summary: Background: Encephalitis is a worldwide public health issue, with a substantially high burden among children in southeast Asia. We aimed to determine the causes of encephalitis in children admitted to hospitals across the Greater Mekong region by implementing a comprehensive state-of-the-art diagnostic procedure harmonised across all centres, and identifying clinical characteristics related to patients’ conditions. Methods: In this multicentre, observational, prospective study of childhood encephalitis, four referral hospitals in Cambodia, Vietnam, Laos, and Myanmar recruited children (aged 28 days to 16 years) who presented with altered mental status lasting more than 24 h and two of the following minor criteria: fever (within the 72 h before or after presentation), one or more generalised or partial seizures (excluding febrile seizures), a new-onset focal neurological deficit, cerebrospinal fluid (CSF) white blood cell count of 5 per mL or higher, or brain imaging (CT or MRI) suggestive of lesions of encephalitis. Comprehensive diagnostic procedures were harmonised across all centres, with first-line testing was done on samples taken at inclusion and results delivered within 24 h of inclusion for main treatable causes of disease and second-line testing was done thereafter for mostly non-treatable causes. An independent expert medical panel reviewed the charts and attribution of causes of all the included children. Using multivariate analyses, we assessed risk factors associated with unfavourable outcomes (ie, severe neurological sequelae and death) at discharge using data from baseline and day 2 after inclusion. This study is registered with ClinicalTrials.gov, NCT04089436, and is now complete. Findings: Between July 28, 2014, and Dec 31, 2017, 664 children with encephalitis were enrolled. Median age was 4·3 years (1·8–8·8), 295 (44%) children were female, and 369 (56%) were male. A confirmed or probable cause of encephalitis was identified in 425 (64%) patients: 216 (33%) of 664 cases were due to Japanese encephalitis virus, 27 (4%) were due to dengue virus, 26 (4%) were due to influenza virus, 24 (4%) were due to herpes simplex virus 1, 18 (3%) were due to Mycobacterium tuberculosis, 17 (3%) were due to Streptococcus pneumoniae, 17 (3%) were due to enterovirus A71, 74 (9%) were due to other pathogens, and six (1%) were due to autoimmune encephalitis. Diagnosis was made within 24 h of admission to hospital for 83 (13%) of 664 children. 119 (18%) children had treatable conditions and 276 (42%) had conditions that could have been preventable by vaccination. At time of discharge, 153 (23%) of 664 children had severe neurological sequelae and 83 (13%) had died. In multivariate analyses, risk factors for unfavourable outcome were diagnosis of M tuberculosis infection upon admission (odds ratio 3·23 [95% CI 1·04–10·03]), coma on day 2 (2·90 [1·78–4·72]), supplementary oxygen requirement (1·89 [1·25–2·86]), and more than 1 week duration between symptom onset and admission to hospital (3·03 [1·68–5·48]). At 1 year after inclusion, of 432 children who were discharged alive from hospital with follow-up data, 24 (5%) had died, 129 (30%) had neurological sequelae, and 279 (65%) had completely recovered. Interpretation: In southeast Asia, most causes of childhood encephalitis are either preventable or treatable, with Japanese encephalitis virus being the most common cause. We provide crucial information that could guide public health policy to improve diagnostic, vaccination, and early therapeutic guidelines on childhood encephalitis in the Greater Mekong region. Funding: Institut Pasteur, Institut Pasteur International Network, Fondation Merieux, Aviesan Sud, INSERM, Wellcome Trust, Institut de Recherche pour le Développement (IRD), and Fondation Total.

Searches for lepton number violating K+→π−e+e+ and K+→π−π0e+e+ decays have been performed using the complete dataset collected by the NA62 experiment at CERN in 2016–2018. Upper limits of 5.3×10−11 and 8.5×10−10 are obtained on the decay branching fractions at 90% confidence level. The former result improves by a factor of four over the previous best limit, while the latter result represents the first limit on the K+→π−π0e+e+ decay rate.

Background: In Marseille, France, following a first severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak in March–May 2020, a second epidemic phase occurred from June, involving 10 new variants. The Marseille-4 variant caused an epidemic that started in August and is still ongoing. Methods: The 1038 SARS-CoV-2 whole genome sequences obtained in our laboratory by next-generation sequencing with Illumina technology were analysed using Nextclade and nextstrain/ncov pipelines and IQ-TREE. A Marseille-4-specific qPCR assay was implemented. Demographic and clinical features were compared between patients with the Marseille-4 variant and those with earlier strains. Results: Marseille-4 harbours 13 hallmark mutations. One leads to an S477N substitution in the receptor binding domain of the spike protein targeted by current vaccines. Using a specific qPCR, it was observed that Marseille-4 caused 12–100% of SARS-CoV-2 infections in Marseille from September 2020, being involved in 2106 diagnoses. This variant was more frequently associated with hypoxemia than were clade 20A strains before May 2020. It caused a re-infection in 11 patients diagnosed with different SARS-CoV-2 strains before June 2020, suggesting either short-term protective immunity or a lack of cross-immunity. Conclusions: Marseille-4 should be considered as a major SARS-CoV-2 variant. Its sudden appearance points towards an animal reservoir, possibly mink. The protective role of past exposure and current vaccines against this variant should be evaluated.

Adeno-associated virus (AAV) vectors are considered efficient vectors for gene transfer, as illustrated by recent successful clinical trials targeting retinal or neurodegenerative disorders. However, limitations as host immune responses to AAV capsid or transduction of limited regions must still be overcome. Here, we focused on locoregional (LR) intravenous perfusion vector delivery that allows transduction of large muscular areas and is considered to be less immunogenic than intramuscular (IM) injection. To confirm this hypothesis, we injected 6 cynomolgus monkeys with an AAV serotype 8 (AAV8) vector encoding for the highly immunogenic GFP driven by either a muscle-specific promoter (n = 3) or a cytomegalovirus (CMV) promoter (n = 3). We report that LR delivery allows long-term GFP expression in the perfused limb (up to 1 year) despite the initiation of a peripheral transgene-specific immune response. The analysis of the immune status of the perfused limb shows that LR delivery induces persisting inflammation. However, this inflammation is not sufficient to result in transgene clearance and is balanced by resident regulatory T cells. Overall, our results suggest that LR delivery promotes persisting transgene expression by induction of Treg cells in situ and might be a safe alternative to IM route to target large muscle territories for the expression of secreted therapeutic factors.

Although SARS-CoV-2 is considered a lung-tropic virus, severe COVID-19 is not just a viral pulmonary infection, clinically it is a multi-organ pathology with major coagulation abnormalities and thromboembolism events. Recently, antiphospholipid (aPL) antibodies were found increased in a large number of COVID-19 patients. Elevated aPL have been well documented in antiphospholipid syndrome (APS), a systemic autoimmune disorder characterized by recurrent venous or arterial thrombosis and/or obstetrical morbidity. Among treatment regimen of APS, hydroxychloroquine (HCQ) is one of the molecules proposed in the primary prevention of thrombosis and obstetrical morbidity in those patients. Due to its antithrombotic properties documented in APS therapy, HCQ could be considered a good candidate for the prevention of thrombotic events in COVID-19 patients in association with anticoagulant and its repurposing deserves further evaluation.

Background & Aims: Direct-acting antiviral (DAA) regimens provide a cure in >95% of patients with chronic HCV infection. However, in some patients in whom therapy fails, resistance-associated substitutions (RASs) can develop, limiting retreatment options and risking onward resistant virus transmission. In this study, we evaluated RAS prevalence and distribution, including novel NS5A RASs and clinical factors associated with RAS selection, among patients who experienced DAA treatment failure. Methods: SHARED is an international consortium of clinicians and scientists studying HCV drug resistance. HCV sequence linked metadata from 3,355 patients were collected from 22 countries. NS3, NS5A, and NS5B RASs in virologic failures, including novel NS5A substitutions, were examined. Associations of clinical and demographic characteristics with RAS selection were investigated. Results: The frequency of RASs increased from its natural prevalence following DAA exposure: 37% to 60% in NS3, 29% to 80% in NS5A, 15% to 22% in NS5B for sofosbuvir, and 24% to 37% in NS5B for dasabuvir. Among 730 virologic failures, most were treated with first-generation DAAs, 94% had drug resistance in ≥1 DAA class: 31% single-class resistance, 42% dual-class resistance (predominantly against protease and NS5A inhibitors), and 21% triple-class resistance. Distinct patterns containing ≥2 highly resistant RASs were common. New potential NS5A RASs and adaptive changes were identified in genotypes 1a, 3, and 4. Following DAA failure, RAS selection was more frequent in older people with cirrhosis and those infected with genotypes 1b and 4. Conclusions: Drug resistance in HCV is frequent after DAA treatment failure. Previously unrecognized substitutions continue to emerge and remain uncharacterized. Lay summary: Although direct-acting antiviral medications effectively cure hepatitis C in most patients, sometimes treatment selects for resistant viruses, causing antiviral drugs to be either ineffective or only partially effective. Multidrug resistance is common in patients for whom DAA treatment fails. Older patients and patients with advanced liver diseases are more likely to select drug-resistant viruses. Collective efforts from international communities and governments are needed to develop an optimal approach to managing drug resistance and preventing the transmission of resistant viruses.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has emerged in Chinese people in December 2019 and has currently spread worldwide causing the COVID-19 pandemic with more than 150,000 deaths. In order for a SARS-CoV like virus circulating in wild life for a very long time to infect the index case-patient, a number of conditions must be met, foremost among which is the encounter with humans and the presence in homo sapiens of a cellular receptor allowing the virus to bind. Recently it was shown that the SARS-CoV-2 spike protein, binds to the human angiotensin I converting enzyme 2 (ACE2). This molecule is a peptidase expressed at the surface of lung epithelial cells and other tissues, that regulates the renin-angiotensin-aldosterone system. Humans are not equal with respect to the expression levels of the cellular ACE2. Moreover, ACE2 polymorphisms were recently described in human populations. Here we review the most recent evidence that ACE2 expression and/or polymorphism could influence both the susceptibility of people to SARS-CoV-2 infection and the outcome of the COVID-19 disease. Further exploration of the relationship between the virus, the peptidase function of ACE2 and the levels of angiotensin II in SARS-CoV-2 infected patients should help to better understand the pathophysiology of the disease and the multi-organ failures observed in severe COVID-19 cases, particularly heart failure.

MacLean and colleagues recently published a very elegant analysis demonstrating that SARS-CoV-2 carries signs of positive selection and that it was already adapted to humans prior to the emergence of COVID-19. Using the Spillover theory as a reference model for zoonotic emergence, they conclude that SARS-CoV-2 must have acquired this human adaptation in bats. We reinterpreted the data from MacLean et al. using a different model of zoonotic emergence as reference, the Circulation model. The use of the Circulation model provides a more parsimonious interpretation showing that this adaptation to humans occurs in the human population after primo infection.

The NA62 experiment at the CERN SPS reports the first search for K+→π+νν¯ using the decay-in-flight technique, based on a sample of 1.21×1011 K+ decays collected in 2016. The single event sensitivity is 3.15×10−10, corresponding to 0.267 Standard Model events. One signal candidate is observed while the expected background is 0.152 events. This leads to an upper limit of 14×10−10 on the K+→π+νν¯ branching ratio at 95% CL.

The NA48/2 experiment at CERN reports the first observation of the K±→π±π0e+e− decay from an exposure of 1.7×1011 charged kaon decays recorded in 2003–2004. A sample of 4919 candidates with 4.9% background contamination allows the determination of the branching ratio in the full kinematic region, BR(K±→π±π0e+e−)=(4.24±0.14)×10−6. The study of the kinematic space shows evidence for a structure dependent contribution in agreement with predictions based on chiral perturbation theory. Several P- and CP-violating asymmetries are also evaluated.

The NA62 experiment at CERN reports searches for K+→μ+N and K+→μ+νX decays, where N and X are massive invisible particles, using the 2016–2018 data set. The N particle is assumed to be a heavy neutral lepton, and the results are expressed as upper limits of O(10−8) of the neutrino mixing parameter |Uμ4|2 for N masses in the range 200–384 MeV/c2 and lifetime exceeding 50 ns. The X particle is considered a scalar or vector hidden sector mediator decaying to an invisible final state, and upper limits of the decay branching fraction for X masses in the range 10–370 MeV/c2 are reported for the first time, ranging from O(10−5) to O(10−7). An improved upper limit of 1.0×10−6 is established at 90% CL on the K+→μ+ννν¯ branching fraction.

Summary: CD32 has raised conflicting results as a putative marker of the HIV-1 reservoir. We measured CD32 expression in tissues from viremic and virally suppressed humanized mice treated relatively early or late after HIV-1 infection with combined antiretroviral therapy. CD32 was expressed in a small fraction of the memory CD4+ T-cell subsets from different tissues in viremic and aviremic mice, regardless of treatment initiation time. CD32+ memory CD4+ T cells were enriched in cell-associated (CA) HIV-1 DNA but not in CA HIV-1 RNA as compared to the CD32−CD4+ fraction. Using multidimensional reduction analysis, several memory CD4+CD32+ T-cell clusters were identified expressing HLA-DR, TIGIT, or PD-1. Importantly, although tissue-resident CD32+CD4+ memory cells were enriched with translation-competent reservoirs, most of it was detected in memory CD32-CD4+ T cells. Our findings support that CD32 labels highly activated/exhausted memory CD4+ T-cell subsets that contain only a small proportion of the translation-competent reservoir.

At the end of November 2019, a novel coronavirus responsible for respiratory tract infections emerged in China. Despite drastic containment measures, this virus, known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spread in Asia and Europe. The pandemic is ongoing with a particular hotspot in southern Europe and America in spring 2020. Many studies predicted an epidemic in Africa similar to that currently seen in Europe and the USA. However, reported data do not confirm these predictions. Several hypotheses that could explain the later emergence and spread of the coronavirus disease 2019 (COVID-19) pandemic in African countries are being discussed, including the lack of health-care infrastructure capable of clinically detecting and confirming COVID-19 cases, the implementation of social distancing and hygiene, international air traffic flows, the climate, the relatively young and rural population, the genetic polymorphism of the angiotensin-converting enzyme 2 receptor, cross-immunity and the use of antimalarial drugs.

The Danish Government announced the culling of 17 million minks in rearing after the report of mink-specific mutations of severe acute respiratory syndrome coronavirus 2 in humans. The rationale behind this decision is that these mutations might negatively impact the deployment of anti-coronavirus disease 2019 vaccines. Is it a precautionary attitude or a panic-driven overreaction?

A search for heavy neutral lepton (N) production in K+→e+N decays using the data sample collected by the NA62 experiment at CERN in 2017–2018 is reported. Upper limits of the extended neutrino mixing matrix element |Ue4|2 are established at the level of 10−9 over most of the accessible heavy neutral lepton mass range 144–462 MeV/c2, with the assumption that the lifetime exceeds 50 ns. These limits improve significantly upon those of previous production and decay searches. The |Ue4|2 range favoured by Big Bang Nucleosynthesis is excluded up to a mass of about 340 MeV/c2.

VortexFitting is a fluid mechanics post-processing tool developed in Python. It aims to detect the presence of vortices in a flow and evaluate their properties. Data obtained from both numerical simulations and experimental flow imaging techniques can be used as inputs. The software supports a number of input file formats such as NetCDF, HD5, TecPlot, and raw text files. The first stage of the vortex search procedure, which is identification of vortex candidates, is accomplished using a set of detection methods: swirling strength, Q criterion, and Δ criterion. The candidate vortices are then fitted to a Lamb–Oseen vortex model using a non-linear least-squares method, and the correlation between the model and the original velocity field is evaluated. If the correlation is deemed high enough, based on a user defined threshold, the vortex is accepted, and properties such as vortex radius and circulation, and vortex center are obtained. Each vortex can be tracked in a transient flow, and its trajectory is reconstructed with its decay characteristics. Two applications are presented in this paper: (i) an experimental columnar vortex moving through a free-surface water channel, and (ii) a numerical simulation of a bubbling fluidized bed. We demonstrate that VortexFitting can successfully identify the presence of vortices and characterize their features in both applications.

Background: A significant proportion of patients develop heart failure (HF) after acute myocardial infarction (MI). Predicting this development with novel biomarkers would allow tailoring healthcare to each individual. We recently identified a circular RNA called MICRA which was associated with HF development after MI. Here, we tested whether MICRA was able to risk stratify MI patients. Methods: MICRA was assessed in whole blood samples collected at reperfusion in 472 patients with acute MI. Left ventricular ejection fraction (EF) was evaluated by echocardiography at 4 months. Multivariable analyses with ordinal regression were conducted to determine the ability of MICRA to classify patients into 3 EF groups: reduced EF (≤40%), mid-range EF (4149%) and preserved EF (≥50%). Results: Eighty seven patients (18%) had a reduced EF, 106 (22%) had a mid-range EF and 279 (59%) had a preserved EF at 4 months. MICRA classified patients into EF groups with an adjusted odds ratio [95% confidence interval] of 0.78 [0.64–0.95]. MICRA improved the predictive value of a multivariable clinical model as attested by a decrease of the Akaike Information Criteria (p = 0.012). Bootstrap internal validation confirmed the incremental prognostic value of MICRA. Conclusion: We report that the circRNA MICRA improves risk classification after MI, supporting the added value of this novel biomarker in future prognostication strategies.

The popularization of genome-wide analyses and RNA sequencing led to the discovery that a large part of the human genome, while effectively transcribed, does not encode proteins. Long non-coding RNAs have emerged as critical regulators of gene expression in both normal and disease states. Studies of long non-coding RNAs expressed in the heart, in combination with gene association studies, revealed that these molecules are regulated during cardiovascular development and disease. Some long non-coding RNAs have been functionally implicated in cardiac pathophysiology and constitute potential therapeutic targets. Here, we review the current knowledge of the function of long non-coding RNAs in the cardiovascular system, with an emphasis on cardiovascular development and biology, focusing on hypertension, coronary artery disease, myocardial infarction, ischemia, and heart failure. We discuss potential therapeutic implications and the challenges of long non-coding RNA research, with directions for future research and translational focus. Keywords: transcriptomics, RNAs, long non-coding RNAs, cardiovascular system, cardiovascular development, cardiovascular disease, vascular disease, therapy, non-coding RNAs

In this paper we discuss results from the study of the energy balance in JET based on calculated heating energies, radiated energy from bolometry and tile calorimetry. Recent data enables us to be more confident in the numbers used and to exclude certain possibilities but the overall energy imbalance which typically amounts to 25% of total input remains unexplained. This shows that caution is required in interpreting fractional radiated powers which are commonly used to measure the effectiveness of impurity seeded scenarios at reducing divertor heat load. Keywords: ILW, JET

In fusion devices, radiofrequency (RF) antennas are used for heating the plasma. Those antennas and the plasma interact with each other through the so-called RF sheaths, layers of plasma where the quasi-neutrality breaks down and large electric fields arise. Among the effects of RF sheaths, there is the enhancement of the particles and energy fluxes toward the surface of the antenna, which in turn generate hot spots and release impurities, which are both deleterious for plasma operations. RF sheaths comprehension stumbles on the difficulty to achieve in situ measurements of the sheath properties, as scrape-off layer plasmas are a harsh environment. The very goal of the ALINE device is to tackle this issue and to fulfil the blank between numerical simulations and full-scale experiment by providing measurements within the RF sheaths in a controlled environment. In this paper we report on the latest experimental results from ALINE, in which a cylindrical Langmuir probe mounted on a remotely controlled and programmable arm allows for plasma characterizations in the three dimensions of space around the stainless steel antenna, including the sheath. We present a series of density and potential profiles and three dimension (3D) maps in the plasma surrounding a stainless-steel RF antenna as well as in the sheath itself, for unmagnetized and magnetized plasmas. Keywords: Magnetized plasma, Sheath, Langmuir probe, Radio-Frequency

Future tokamak reactors of conventional design will require high levels of exhaust power dissipation (more than 90% of the input power) if power densities at the divertor targets are to remain compatible with active cooling. Impurity seeded H-mode discharges in JET-ITER-like Wall (ILW) have reached a maximum radiative fraction (Frad) of ∼75%. Divertor Langmuir probe (LP) measurements in these discharges indicate, however, that less than ∼3% of the thermal plasma power reaches the targets, suggesting a missing channel for power loss. This paper presents experimental evidence from limiter LP for enhanced cross-field particle fluxes on the main chamber walls at high Frad. In H-mode nitrogen-seeded discharges with Frad increasing from ∼30% to up to ∼75%, the main chamber wall particle fluence rises by a factor ∼3 while the divertor plasma fluence drops by one order of magnitude. Contribution of main chamber wall particle losses to detachment, as suggested by EDGE2D-EIRENE modeling, is not sufficient to explain the magnitude of the observed divertor fluence reduction. An intermediate detached case obtained at Frad ∼ 60% with neon seeding is also presented. Heat loads were measured using the main chamber wall thermocouples. Comparison between thermocouple and bolometry measurements shows that the fraction of the input power transported to the main chamber wall remains below ∼5%, whatever the divertor detachment state is. Main chamber sputtering of beryllium by deuterium is reduced in detached conditions only on the low field side. If the fraction of power exhaust dissipated to the main chamber wall by cross-field transport in future reactors is similar to the JET-ILW levels, wall plasma power loading should not be an issue. However, other contributions such as charge exchange may be a problem.

The World Health Organization (WHO) has declared that hepatitis C virus (HCV) should be eliminated as a public health threat. A key recommendation to reach this elimination goal, is to reduce new infections by 90% and liver-related mortality by 65%. Highly effective direct-acting antiviral agents (DAA) play a major role in this elimination. Unfortunately, DAA treatment fails approximately 2.5–5% of patients, often in the presence of resistance-associated substitutions (RAS). This could eventually lead to a total number of 1.8–3.6 million first-line DAA failures. RAS may jeopardise the elimination goals for several reasons; most importantly, virus transmission and infection progression will continue. More data are required to handle RAS adequately and identify mutational patterns causing resistance. Currently, sample sizes are small, data are scattered and methods heterogenic. Collaboration is therefore key and a European collaboration, such as HEPCARE, should provide a solution.

Linear friction welding (LFW) is a near net shape solid state joining technology for aerospace applications. In this study LFW of a recently introduced Ni-based superalloy, AD730™, with superior properties for use in the hot section of gas turbines was studied. In order to minimize the number of experiments for achieving sound welds, an analytical method was developed that allowed to determine the optimum process parameters with a limited number of experiments. The predictions of the method were validated by LFW experiments, and sound samples without defects were produced. Microstructure evolution of the as-welded samples from the weld center to the base metal was investigated using laser confocal and field emission scanning electron microscopy (SEM) including electron back scatter diffraction (EBSD), energy dispersive spectrometer (EDS) and backscattered imaging (BSE). Post-weld heat treatment (PWHT), consisting of a γ′ sub-solvus solutionizing followed by aging, was conducted on LFWed blocks. Then, microstructure evolution and mechanical testing were conducted on PWHTed samples and compared with the as welded blocks. The obtained results were correlated with microhardness as well as tensile testing at room temperature and 650 °C, and interpreted in terms of fundamental metallurgical processes. Macroscopic examination of the PWHTed joints revealed that the samples failed out of the weld zone, further demonstrating the appropriate selection of LFW processing parameters using the proposed analytical method. PWHTed samples exhibited better room and high temperature tensile properties compared to those of the as-welded samples. Microscopic examination of the fracture zones of the samples showed that the higher levels of reprecipitation of γ′ particles in the thermomechanically-affected zone (TMAZ) of the PWHT samples were associated to their higher tensile properties compared to the as welded ones. Keywords: AD730™ Ni-base superalloy, Linear friction welding, Process parameter, Analytical method, Microstructure, Mechanical property

The NA62 experiment at CERN reports a search for the lepton number violating decays K+→π−e+e+ and K+→π−μ+μ+ using a data sample collected in 2017. No signals are observed, and upper limits on the branching fractions of these decays of 2.2×10−10 and 4.2×10−11 are obtained, respectively, at 90% confidence level. These upper limits improve on previously reported measurements by factors of 3 and 2, respectively.

The Institut Hospitalo-Universitaire Méditerranée Infection is positioned for the diagnosis, prevention and treatment of the ‘big three’ killer diseases: malaria, tuberculosis and HIV. We implemented the use of new diagnostic samples such as stools and new diagnostic tests such as mass spectrometry for the dual identification of vectors and pathogens. Furthermore, advances in the prevention and treatment of malaria and tuberculosis are reviewed, along with advances in the understanding of the role of microbiota in the resistance to HIV infection. These achievements represent a major step towards a better management of the ‘big three’ diseases worldwide. Keywords: Diagnostic, drugs, HIV, malaria, microbiota, Mycobacterium tuberculosis, Plasmodium falciparum, resistance, tuberculosis, vector