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Abstract The emergence of the Middle Palaeolithic, and its variability over time and space are key questions in the field of prehistoric archaeology. Many sites have been documented in the south-eastern margins of the Massif central and the middle Rhône valley, a migration path that connects Northern Europe with the Mediterranean. Well-dated, long stratigraphic sequences are essential to understand Neanderthals dynamics and demise, and potential interactions with Homo sapiens in the area, such as the one displayed at the Maras rock shelter (“Abri du Maras”). The site is characterised by exceptional preservation of archaeological remains, including bones dated using radiocarbon (14C) and teeth using electron spin resonance combined with uranium series (ESR/U-series). Optically stimulated luminescence was used to date the sedimentary deposits. By combining the new ages with previous ones using Bayesian modelling, we are able to clarify the occupation time over a period spanning 200,000 years. Between ca. 250 and 40 ka, the site has been used as a long-term residence by Neanderthals, specifically during three interglacial periods: first during marine isotopic stage (MIS) 7, between 247 ± 34 and 223 ± 33 ka, and then recurrently during MIS 5 (between 127 ± 17 and 90 ± 9 ka) and MIS 3 (up to 39,280 cal BP).

Abstract Motorized spiral enteroscopy (MSE) enhances small bowel exploration, but the optimal target population for this technique is unknown. We aimed to identify the target population for MSE by evaluating its efficacy and safety, as well as detecting predictors of efficacy. A prospective multicenter observational study was conducted at 9 tertiary hospitals in Spain, enrolling patients between June 2020–2022. Analyzed data included demographics, indications for the procedure, exploration time, depth of maximum insertion (DMI), technical success, diagnostic yield, interventional yield, and adverse events (AE) up to 14 days from enteroscopy. Patients with prior gastrointestinal surgery, unsuccessful balloon enteroscopy and small bowel strictures were analyzed. A total of 326 enteroscopies (66.6% oral route) were performed in 294 patients (55.1% males, 65 years ± 21). Prior abdominal surgery was present in 50% of procedures (13.5% gastrointestinal surgery). Lower DMI (162 vs 275 cm, p = 0.037) and diagnostic yield (47.7 vs 67.5%, p = 0.016) were observed in patients with prior gastrointestinal surgery. MSE showed 92.2% technical success and 56.9% diagnostic yield after unsuccessful balloon enteroscopy (n = 51). In suspected small bowel strictures (n = 49), the finding was confirmed in 23 procedures (46.9%). The total AE rate was 10.7% (1.8% classified as major events) with no differences related to prior gastrointestinal/abdominal surgery, unsuccessful enteroscopy, or suspected small bowel strictures. The study demonstrates that MSE has a lower diagnostic yield and DMI in patients with prior gastrointestinal surgery but is feasible after unsuccessful balloon-enteroscopy and in suspected small bowel strictures without safety concerns.

Objetives. Bacterial resistance to antibiotics is a public health problem, however, few studies are performed in natural water ecosystems in middle-low-income countries. The objective of this study was to evaluate the presence and sensitivity to antimicrobials of Escherichia coli strains isolated from 24 irrigation water samples from the Rimac River in eastern Lima. Materials and methods. E.coli were identified by PCR. Antibiotic susceptibility was determined by disk diffusion method. The extended-spectrum beta-lactamase (ESBL), quinolones and virulence involved genes were determined by PCR. Results. All samples exceeded the permissible limits established in the Environmental Quality Standards for vegetable irrigation. Of the 94 strains, 72.3% showed resistance to at least one antibiotic, 24.5% were multidrug-resistant (MDR), and 2.1% were extremely drug resistant. The highest percentages of resistance were observed against nalidixic acid (50%), trimethoprim-sulfamethoxazole (35.5%) and ciprofloxacin (20.4%). Among the isolates, 3.2% presented ESBL phenotype related to blaCTX-M-15 gene. The transferable mechanisms of resistance to quinolones, qnrB were more frequent (20.4%), and 2.04% had the qnrS. It was determined that 5.3% were diarrheagenic E. coli and of these, 60% were enterotoxigenic E. coli, 20% were enteropathogenic E. coli and 20% were enteroaggregative E. coli. Conclusions. Our results show the existence diarrheagenic pathotypes in water used to irrigate fresh produce and highlights the presence of ESBL-producers and MDR E. coli, demonstrating the role that irrigation water plays in disseminating resistance genes in Peru.

Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity.

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.

Objectives To compare the treatment effectiveness of secukinumab in radiographic (r) versus non-radiographic (nr) axial spondyloarthritis (axSpA) patients treated in routine care across Europe.Methods Prospectively collected data on secukinumab-treated axSpA patients with known radiographic status were pooled from nine countries.Remission rates based on patient-reported outcomes (PROs; Numeric Rating Scale (0–10), for example, pain ≤2/Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) ≤2 and Ankylosing Spondylitis Disease Activity Score (ASDAS) inactive disease (ID)

Introduction Lymphoedema is a chronic condition caused by lymphatic insufficiency. It leads to swelling of the limb/midline region and an increased risk of infection. Lymphoedema is often associated with mental and physical problems limiting quality of life. The first choice of treatment is a conservative treatment, consisting of exercises, skin care, lymph drainage and compression. Reconstructive lymphatic surgery is also often performed, that is, lymphovenous anastomoses, lymph node transfer or a combination. However, robust evidence on the effectiveness of reconstructive lymphatic surgery is missing. Therefore, the objective of this trial is to investigate the added value of reconstructive lymphatic surgery to the conservative treatment in patients with lymphoedema.Methods and analysis A multicentre randomised controlled and pragmatic trial was started in March 2022 in three Belgian university hospitals. 90 patients with arm lymphoedema and 90 patients with leg lymphoedema will be included. All patients are randomised between conservative treatment alone (control group) or conservative treatment with reconstructive lymphatic surgery (intervention group). Assessments are performed at baseline and at 1, 3, 6, 12, 18, 24 and 36 months. The primary outcome is lymphoedema-specific quality of life at 18 months. Key secondary outcomes are limb volume and duration of wearing the compression garment at 18 months. The approach of reconstructive lymphatic surgery is based on presurgical investigations including clinical examination, lymphofluoroscopy, lymphoscintigraphy, lymph MRI or CT angiography (if needed). All patients receive conservative treatment during 36 months, which is applied by the patient’s own physical therapist and by the patient self. From months 7 to 12, the hours a day of wearing the compression garment are gradually decreased.Ethics and dissemination The study has been approved by the ethical committees of University Hospitals Leuven, Ghent University Hospital and CHU UCL Namur. Results will be disseminated via peer-reviewed journals and presentations.Trial registration number NCT05064176

Abstract Background Neoantigens are patient- and tumor-specific peptides that arise from somatic mutations. They stand as promising targets for personalized therapeutic cancer vaccines. The identification process for neoantigens has evolved with the use of next-generation sequencing technologies and bioinformatic tools in tumor genomics. However, in-silico strategies for selecting immunogenic neoantigens still have very low accuracy rates, since they mainly focus on predicting peptide binding to Major Histocompatibility Complex (MHC) molecules, which is key but not the sole determinant for immunogenicity. Moreover, the therapeutic potential of neoantigen-based vaccines may be enhanced using an optimal delivery platform that elicits robust de novo immune responses. Methods We developed a novel neoantigen selection pipeline based on existing software combined with a novel prediction method, the Neoantigen Optimization Algorithm (NOAH), which takes into account structural features of the peptide/MHC-I interaction, as well as the interaction between the peptide/MHC-I complex and the TCR, in its prediction strategy. Moreover, to maximize neoantigens’ therapeutic potential, neoantigen-based vaccines should be manufactured in an optimal delivery platform that elicits robust de novo immune responses and bypasses central and peripheral tolerance. Results We generated a highly immunogenic vaccine platform based on engineered HIV-1 Gag-based Virus-Like Particles (VLPs) expressing a high copy number of each in silico selected neoantigen. We tested different neoantigen-loaded VLPs (neoVLPs) in a B16-F10 melanoma mouse model to evaluate their capability to generate new immunogenic specificities. NeoVLPs were used in in vivo immunogenicity and tumor challenge experiments. Conclusions Our results indicate the relevance of incorporating other immunogenic determinants beyond the binding of neoantigens to MHC-I. Thus, neoVLPs loaded with neoantigens enhancing the interaction with the TCR can promote the generation of de novo antitumor-specific immune responses, resulting in a delay in tumor growth. Vaccination with the neoVLP platform is a robust alternative to current therapeutic vaccine approaches and a promising candidate for future personalized immunotherapy.

Abstract Novel perioperative strategies are needed to reduce recurrence rates in patients undergoing nephrectomy for high-risk, non-metastatic clear cell renal cell carcinoma (ccRCC). We conducted a prospective, phase I trial of neoadjuvant nivolumab prior to nephrectomy in 15 evaluable patients with non-metastatic ccRCC. We leveraged tissue from that cohort to elucidate the effects of PD-1 inhibition on immune cell populations in ccRCC and correlate the evolving immune milieu with anti-PD-1 response. We found that nivolumab durably induces a pro-inflammatory state within the primary tumor, and baseline immune infiltration within the primary tumor correlates with nivolumab responsiveness. Nivolumab increases CTLA-4 expression in the primary tumor, and subsequent nephrectomy increases circulating concentrations of sPD-L1, sPD-L3 (sB7-H3), and s4-1BB. These findings form the basis to consider neoadjuvant immune checkpoint inhibition (ICI) for high-risk ccRCC while the tumor remains in situ and provide the rationale for perioperative strategies of novel ICI combinations.

El objetivo de esta carta es poner en evidencia la realidad a la que puede exponerse el especialista en Reumatología en un contexto diferente. El Grupo Latinoamericano de Estudios de Enfermedades Reumáticas en Pueblos Originarios (GLADERPO) trabaja, desde su conformación en 2009, en diferentes comunidades de Latinoamérica. En Argentina, en las comunidades Qom de las ciudades de Rosario (Santa Fe) y Presidencia Roque Sáenz Peña (Chaco), y en la comunidad Wichí de la localidad “Misión Chaqueña, El Algarrobal” (Salta). Los objetivos fundamentales de este proyecto son, por un lado, estimar la prevalencia de enfermedades reumáticas y, por el otro, describir la percepción y las barreras en el acceso al sistema de salud. Un punto central del mismo ha sido la intervención en la comunidad a través del seguimiento de pacientes detectados con enfermedades reumáticas, principalmente artritis reumatoide (AR), y el diseño e implementación de diferentes estrategias educativas, con la intención de visibilizar la dificultad en el acceso al sistema de salud de estas comunidades, intentando disminuir la brecha existente.

Background: Identifying potential factors correlated with the sustained presence of antibodies in plasma may facilitate improved retrospective diagnoses and aid in the appraisal of pertinent vaccination strategies for various demographic groups. The main objective was to describe the persistence of anti-spike IgG one year after diagnosis in children and analyse its levels in relation to epidemiological and clinical variables. Methods: A prospective, longitudinal, observational study was conducted in a university reference hospital in the Metropolitan Region of Barcelona (Spain) (March 2020–May 2021). This study included patients under 18 years of age with SARS-CoV-2 infection (positive PCR or antigen tests for SARS-CoV-2). Clinical and serological follow-up one year after infection was performed. Results: We included 102 patients with a median age of 8.8 years. Anti-spike IgG was positive in 98/102 (96%) 12 months after the infection. There were higher anti-spike IgG levels were noted in patients younger than 2 years (p = 0.034) and those with pneumonia (p < 0.001). A positive and significant correlation was observed between C-reactive protein at diagnosis and anti-spike IgG titre one-year after diagnosis (p = 0.027). Conclusion: Anti-SARS-CoV-2 IgG antibodies were detected in almost all paediatric patients one year after infection. We also observed a positive correlation between virus-specific IgG antibody titres with SARS-CoV-2 clinical phenotype (pneumonia) and age (under 2 years old).

Bed bug infestations have been increasing worldwide since the 2000s. Their consequences for health range from skin reactions to major psychological distress. However, epidemiological data is still lacking. This study estimated the incidence of general practice consultations related to bed bugs in France and evaluated factors associated with repercussions on everyday life. We conducted a prospective observational study from March 2019 to April 2020 among 217 GPs in France. Annual and monthly GP consultations related to bed bugs were estimated from the reported cases. Descriptive analyses were performed, and factors associated with repercussions on everyday life were assessed using a logistic regression model. The annual incidence rate of GP consultations related to bed bugs was estimated at 109 per 100,000 inhabitants [95%CI: 92-126]. Bed bug-related consultations occurred in all regions, peaking in the summer. Moderate-to-severe repercussions on everyday life were reported by 39% of the patients. The associated factors with this level of repercussions were: having seen bed bugs (OR = 4.02 [95%CI: 1.6-10.73]), having lesions from scratching (OR = 5.25 [1.65-19.92]), having lesions on the head and the neck (OR = 3.97 [1.52-10.95]) and reporting psychological distress (OR = 6.79 [2.47-20.42]). This study provides new knowledge on GP consultations related to bed bugs in France. These data will help tailor public health programs to the population's needs, including information and training for primary healthcare professionals.

[This corrects the article DOI: 10.1371/journal.pone.0279815.].

Cognitive difficulties are reported as lasting sequelae within post COVID-19 condition. However, the chronicity of these difficulties and related factors of fatigue, mood, and perceived health have yet to be fully determined. To address this, the current longitudinal study aimed to clarify the trends of cognitive test performance and cognitive domain impairment following COVID-19 onset, and whether hospitalization influences outcomes. 57 participants who reported subjective cognitive difficulties after confirmed COVID-19 infection were assessed at baseline (~6 months post COVID-19) and follow-up (~15 months later) visits. Assessments included measures across multiple cognitive domains and self-report questionnaires of fatigue, mood, and overall health. Analyses were conducted in three stages: at the test score level (raw and adjusted scores), at the cognitive domain level, and stratified by hospitalization status during infection. Results at the test-score level indicate that cognitive performance remains relatively stable across assessments at the group level, with no significant improvements in any adjusted test scores at follow-up. Cognitive domain analyses indicate significant reductions in attention and executive functioning impairment, while memory impairment is slower to resolve. On self-report measures, there was a significant improvement in overall health ratings at follow-up. Finally, those hospitalized during infection performed worse on timed cognitive measures across visits and accounted for a larger proportion of cases with short-term and working memory impairment at follow-up. Overall, our findings indicate that cognitive difficulties persist both at test score and cognitive domain levels in many cases of post COVID-19 condition, but evidence suggests some improvement in global measures of attention, executive functioning and overall self-rated health. Furthermore, an effect of hospitalization on cognitive symptoms post COVID-19 may be more discernible over time.

Health facility delivery is one of the critical indicators to monitor progress towards the provision of skilled delivery care and reduction in perinatal mortality. In Ethiopia, utilization of health facilities for skilled delivery care has been increasing but varies greatly by region and among specific socio-demography groups. We aimed to measure the prevalence and determinants of health facility delivery in the Amhara region in Ethiopia. From December 2018 to November 2020, we conducted a longitudinal study from a cohort of 2801 pregnant women and described the location of delivery and the association with determinants. We interviewed a subset of women who delivered in the community and analyzed responses using the three delays model to understand reasons for not using health facility services. A multivariable poisson regression model with robust error variance was used to estimate the presence and magnitude of association between location of delivery and the determinants. Of the 2,482 pregnant women followed through to birth, 73.6% (n = 1,826) gave birth in health facilities, 24.3% (n = 604) gave birth at home and 2.1% (n = 52) delivered on the way to a health facility. Determinants associated with increased likelihood of delivery at a health facility included formal maternal education, shorter travel times to health facilities, primiparity, higher wealth index and having attended at least one ANC visit. Most common reasons mothers gave for not delivering in a health facility were delays in individual/family decision to seek care. The proportion of deliveries occurring in health facilities is increasing but falls below targets. Interventions that focus on the identified social-demographic determinants and delays are warranted.

Summary: Background: Diagnosing Cushing’s syndrome (CS) is highly complex. As the diagnostic potential of urinary steroid metabolome analysis by gas chromatography-mass spectrometry (GC-MS) in combination with systems biology has not yet been fully exploited, we studied a large cohort of patients with CS. Methods: We quantified daily urinary excretion rates of 36 steroid hormone metabolites. Applying cluster analysis, we investigated a control group and 168 patients: 44 with Cushing’s disease (CD) (70% female), 18 with unilateral cortisol-producing adrenal adenoma (83% female), 13 with primary bilateral macronodular adrenal hyperplasia (PBMAH) (77% female), and 93 ruled-out CS (73% female). Findings: Cluster-Analysis delineated five urinary steroid metabotypes in CS. Metabotypes 1, 2 and 3 revealing average levels of cortisol and adrenal androgen metabolites included patients with exclusion of CS or and healthy controls. Metabotype 4 reflecting moderately elevated cortisol metabolites but decreased DHEA metabolites characterized the patients with unilateral adrenal CS and PBMAH. Metabotype 5 showing strong increases both in cortisol and DHEA metabolites, as well as overloaded enzymes of cortisol inactivation, was characteristic of CD patients. 11-oxygenated androgens were elevated in all patients with CS. The biomarkers THS, F, THF/THE, and (An + Et)/(11β-OH-An + 11β-OH-Et) correctly classified 97% of patients with CS and 95% of those without CS. An inverse relationship between 11-deoxygenated and 11-oxygenated androgens was typical for the ACTH independent (adrenal) forms of CS with an accuracy of 95%. Interpretation: GC-MS based urinary steroid metabotyping allows excellent identification of patients with endogenous CS and differentiation of its subtypes. Funding: The study was funded by the Else Kröner-Fresenius-Stiftung and the Eva-Luise-und-Horst-Köhler-Stiftung.

Con el objetivo de describir las características clínicas y epidemiológicas de los pacientes fallecidos por dengue durante el 2017 en Piura, se realizó un estudio retrospectivo de revisión de 24 historias clínicas. El 67% de los casos fueron mujeres y tres (12,5%) estaban embarazadas. La diabetes (12,5%) y la hipertensión (16,7%) fueron las comorbilidades más frecuentes. Sólo en el 12,5% se reportó dengue previo. El tiempo transcurrido desde asistencia sanitaria hasta la muerte fue de 4,10 (DE: 5,34) días. Se hicieron transfusiones de glóbulos rojos en el 45,8% de los casos, plasma en el 25%, plaquetas en el 16,8% y crioprecipitado en el 16,8%. También fueron frecuentes la terapia con ristaloides (91,7%) y el tratamiento con fármacos vasoactivos (70,8%). En conclusión, la mortalidad del dengue grave fue mayoritaria en las mujeres adultas y el tiempo de atención desde el primer establecimiento de salud hasta una unidad especializada fue prolongada.

India reported >10 million coronavirus disease (COVID-19) cases and 149,000 deaths in 2020. To reassess reported deaths and estimate incidence rates during the first 6 months of the epidemic, we used a severe acute respiratory syndrome coronavirus 2 transmission model fit to data from 3 serosurveys in Delhi and time-series documentation of reported deaths. We estimated 48.7% (95% credible interval 22.1%–76.8%) cumulative infection in the population through the end of September 2020. Using an age-adjusted overall infection fatality ratio based on age-specific estimates from mostly high-income countries, we estimated that just 15.0% (95% credible interval 9.3%–34.0%) of COVID-19 deaths had been reported, indicating either substantial underreporting or lower age-specific infection-fatality ratios in India than in high-income countries. Despite the estimated high attack rate, additional epidemic waves occurred in late 2020 and April–May 2021. Future dynamics will depend on the duration of natural and vaccine-induced immunity and their effectiveness against new variants.

Abstract Neurologic impairment persisting months after acute severe SARS-CoV-2 infection has been described because of several pathogenic mechanisms, including persistent systemic inflammation. The objective of this study is to analyze the selective involvement of the different cognitive domains and the existence of related biomarkers. Cross-sectional multicentric study of patients who survived severe infection with SARS-CoV-2 consecutively recruited between 90 and 120 days after hospital discharge. All patients underwent an exhaustive study of cognitive functions as well as plasma determination of pro-inflammatory, neurotrophic factors and light-chain neurofilaments. A principal component analysis extracted the main independent characteristics of the syndrome. 152 patients were recruited. The results of our study preferential involvement of episodic and working memory, executive functions, and attention and relatively less affectation of other cortical functions. In addition, anxiety and depression pictures are constant in our cohort. Several plasma chemokines concentrations were elevated compared with both, a non-SARS-Cov2 infected cohort of neurological outpatients or a control healthy general population. Severe Covid-19 patients can develop an amnesic and dysexecutive syndrome with neuropsychiatric manifestations. We do not know if the deficits detected can persist in the long term and if this can trigger or accelerate the onset of neurodegenerative diseases.

Abstract We study non-relativistic propagation of Gaussian wave packets in one-dimensional Eckart potential, a barrier, or a well. In the picture used, the transmitted wave packet results from interference between the copies of the freely propagating state with different spatial shifts (delays), $$x'$$ x ′ , induced by the scattering potential. The Uncertainty Principle precludes relating the particle’s final position to the delay experienced in the potential, except in the classical limit. Beyond this limit, even defining an effective range of the delay is shown to be an impracticable task, owing to the oscillatory nature of the corresponding amplitude distribution. Our examples include the classically allowed case, semiclassical tunnelling, delays induced in the presence of a virtual state, and scattering by a low barrier. The properties of the amplitude distribution of the delays, and its pole representation are studied in detail.

Abstract Homeostatic renal filtration relies on the integrity of podocytes, which function in glomerular filtration. These highly specialized cells are damaged in 90% of chronic kidney disease, representing the leading cause of end-stage renal failure. Although modest podocyte renewal has been documented in adult mice, the mechanisms regulating this process remain largely unknown and controversial. Using a mouse model of Adriamycin-induced nephropathy, we find that the recovery of filtration function requires up-regulation of the endogenous telomerase component TERT. Previous work has shown that transient overexpression of catalytically inactive TERT (i-TERTci mouse model) has an unexpected role in triggering dramatic podocyte proliferation and renewal. We therefore used this model to conduct specific and stochastic lineage-tracing strategies in combination with high throughput sequencing methods. These experiments provide evidence that TERT drives the activation and clonal expansion of podocyte progenitor cells. Our findings demonstrate that the adult kidney bears intrinsic regenerative capabilities involving the protein component of telomerase, paving the way for innovative research toward the development of chronic kidney disease therapeutics.

The hippocampus is a fundamental cortical structure in the memory process of encoding, retaining, and recalling information. Transcranial Magnetic Stimulation (TMS) following a Paired Associative Stimulation (PAS) enhances nervous system excitability and promotes cortical plasticity mechanisms by synchronizing two stimuli in the same neural pathway. However, PAS has not been shown to improve memorization capacity yet. Here, we present an innovative protocol stemming from the PAS paradigm, which combines single-pulse TMS to the hippocampus with endogenous hippocampal activity during a working memory (WM) task. 96 volunteers were randomized across one experimental group and three parallel groups (motor cortex stimulation, sham stimulation, and no stimulation) in a single session. This combined-stimuli configuration resulted in an increased memorization capacity in the WM task, which was dependent on the stimulated brain location and subjects' basal memory performance. These results are potentially significant for clinical research on memory dysfunction and its related neurological disorders. Future research on paired associative or combined stimulation is required to unveil stimulation-derived neural mechanisms that enhance the ability to memorize.

Objectives Ambulance officers administering methoxyflurane as an inhalational analgesic may be exposed to trace vapor. Fluoride is a methoxyflurane metabolite, and has been associated with acute renal failure in anesthesia patients and skeletal fluorosis with chronic elevated serum levels from other sources. However, there has been no direct measurement of serum fluoride in occupationally exposed ambulance officers. Thus, this study directly measures serum fluoride over a prolonged period in order to determine renal toxic and skeletal fluorosis risk to ambulance officers who are administering methoxyflurane. Material and Methods Serum inorganic fluoride concentrations were measured in a prospective observational study of 12 emergency medical technicians (EMTs). The study took 7 serum fluoride measurements over 24 months. A meta-analysis of healthy adult serum fluoride ranges was also conducted. Results The typical healthy adult serum fluoride range was determined to be 0.21–2.11 μmol/l (p < 0.001). The EMTs’ baseline median (IQR) serum fluoride concentrations were 0.4 μmol/l (0.2; 1.0) with maximum 1.6 μmol/l. The EMTs’ overall median serum fluoride was 0.4 μmol/l (0.2; 1.3) with maximum 4.0 μmol/l, usually within healthy reference ranges. All results were ≤10% of the suggested single-dose renal toxic threshold. One result was above a threshold for skeletal fluorosis. The highest measured serum fluoride was 24% of the lowest level associated with radiologic evidence of fluorosis. There was no evidence overall of increasing serum fluoride levels. Conclusions There was no evidence that EMTs’ exposure to methoxyflurane resulted in sustained increased serum fluoride. These results imply EMTs’ occupational safety from acute renal toxicity when activated carbon filtration is used on patient exhalation. However, 1 serum fluoride result above a skeletal fluorosis threshold suggests that the risk of mild skeletal fluorosis cannot be excluded. Int J Occup Med Environ Health. 2021;34(6):767–77

Abstract The combinatorial study of phylogenetic networks has attracted much attention in recent times. In particular, one class of them, the so-called tree-child networks, are becoming the most prominent ones. However, their combinatorial properties are largely unknown. In this paper we address the problem of exactly counting them. We conjecture a relationship with the cardinality of a certain class of words. By solving the counting problem for the words, and on the basis of the conjecture, several simple recurrence formulas for general cases arise. Moreover, a precise asymptotic analysis is provided. Our results coincide with all current formulas in the literature for particular subclasses of tree-child networks, as well as with numerical results obtained for small networks. We expect that the study of the relationship between the newly defined words and the networks will lead to further combinatoric characterizations of this class of phylogenetic networks.

Abstract Inferring the interactions between coupled oscillators is a significant open problem in complexity science, with multiple interdisciplinary applications. While the Kalman filter (KF) technique is a well-known tool, widely used for data assimilation and parameter estimation, to the best of our knowledge, it has not yet been used for inferring the connectivity of coupled chaotic oscillators. Here we demonstrate that KF allows reconstructing the interaction topology and the coupling strength of a network of mutually coupled Rössler-like chaotic oscillators. We show that the connectivity can be inferred by considering only the observed dynamics of a single variable of the three that define the phase space of each oscillator. We also show that both the coupling strength and the network architecture can be inferred even when the oscillators are close to synchronization. Simulation results are provided to show the effectiveness and applicability of the proposed method.

Abstract Background Surveillance tools to estimate viral transmission dynamics in young populations are essential to guide recommendations for school opening and management during viral epidemics. Ideally, sensitive techniques are required to detect low viral load exposures among asymptomatic children. We aimed to estimate SARS-CoV-2 infection rates in children and adult populations in a school-like environment during the initial COVID-19 pandemic waves using an antibody-based field-deployable and non-invasive approach. Methods Saliva antibody conversion defined as ≥ 4-fold increase in IgM, IgA, and/or IgG levels to five SARS-CoV-2 antigens including spike and nucleocapsid constructs was evaluated in 1509 children and 396 adults by high-throughput Luminex assays in samples collected weekly in 22 summer schools and 2 pre-schools in 27 venues in Barcelona, Spain, from June 29th to July 31st, 2020. Results Saliva antibody conversion between two visits over a 5-week period was 3.22% (49/1518) or 2.36% if accounting for potentially cross-reactive antibodies, six times higher than the cumulative infection rate (0.53%) assessed by weekly saliva RT-PCR screening. IgG conversion was higher in adults (2.94%, 11/374) than children (1.31%, 15/1144) (p=0.035), IgG and IgA levels moderately increased with age, and antibodies were higher in females. Most antibody converters increased both IgG and IgA antibodies but some augmented either IgG or IgA, with a faster decay over time for IgA than IgG. Nucleocapsid rather than spike was the main antigen target. Anti-spike antibodies were significantly higher in individuals not reporting symptoms than symptomatic individuals, suggesting a protective role against COVID-19. Conclusion Saliva antibody profiling including three isotypes and multiplexing antigens is a useful and user-friendlier tool for screening pediatric populations to detect low viral load exposures among children, particularly while they are not vaccinated and vulnerable to highly contagious variants, and to recommend public health policies during pandemics.

Abstract The aim of this study was to analyze the degree of agreement comparing number and distance covered in different acceleration and deceleration sections registered by a video tracking system (MEDIACOACH) and a GPS device (WIMU PRO) during official competition. Data from a Spanish professional club were registered over the course of a season. First, the descriptive statistics presented more bursts of accelerations and decelerations in WIMU PRO than in MEDIACOACH, whereas the distances covered recorded by both systems were similar. Second, negative relationships were found (i.e., negative bias) comparing WIMU PRO to MEDIACOACH in the number of accelerations and decelerations between 0/1 m/s2 and ½ m/s2 (p 0.19) and small (0.2–0.59) for most variables. The standardized typical errors in the estimate (TEE) were moderate (0.3–0.59) and small to moderate (0.1–0.29 to 0.3–0.59), respectively. Also, the Intra class Correlation Coefficients (ICCs) for agreement and consistency between systems showed good and excellent values (> 0.90). The magnitude of change in means (%) between systems, defined as the percentage change between the numbers or values, was below 14% and 7% for number and distances covered, respectively. All scores in the smallest worthwhile change were lower than 9% and in the coefficients of variation were lower than 95% and 15%, respectively. Thus, both systems demonstrated an acceptable degree of agreement and could be useful in analyzing players’ acceleration demands in professional soccer. However, caution is required when interpreting the results and a comparison with a gold standard is required in order to validate both systems.

Abstract The clumped isotope composition (Δ47, the anomaly of the mass 47 isotopologue relative to the abundance expected from a random isotope distribution) of CO2 has been suggested as an additional tracer for gross CO2 fluxes. However, the effect of photosynthetic gas exchange on Δ47 has not been directly determined and two indirect/conceptual studies reported contradicting results. In this study, we quantify the effect of photosynthetic gas exchange on Δ47 of CO2 using leaf cuvette experiments with one C4 and two C3 plants. The experimental results are supported by calculations with a leaf cuvette model. Our results demonstrate the important roles of the Δ47 value of CO2 entering the leaf, kinetic fractionation as CO2 diffuses into, and out of the leaf and CO2–H2O isotope exchange with leaf water. We experimentally confirm the previously suggested dependence of Δ47 of CO2 in the air surrounding a leaf on the stomatal conductance and back-diffusion flux. Gas exchange can enrich or deplete the Δ47 of CO2 depending on the Δ47 of CO2 entering the leaf and the fraction of CO2 exchanged with leaf water and diffused back to the atmosphere, but under typical ambient conditions, it will lead to a decrease in Δ47.

Abstract DNA-Damage Response (DDR) proteins are crucial for maintaining the integrity of the genome by identifying and repairing errors in DNA. Variants affecting their function can have severe consequences since failure to repair damaged DNA can result in cells turning cancerous. Here, we compare germline and somatic variants in DDR genes, specifically looking at their locations in the corresponding three-dimensional (3D) structures, Pfam domains, and protein–protein interaction interfaces. We show that somatic variants in metastatic cases are more likely to be found in Pfam domains and protein interaction interfaces than are pathogenic germline variants or variants of unknown significance (VUS). We also show that there are hotspots in the structures of ATM and BRCA2 proteins where pathogenic germline, and recurrent somatic variants from primary and metastatic tumours, cluster together in 3D. Moreover, in the ATM, BRCA1 and BRCA2 genes from prostate cancer patients, the distributions of germline benign, pathogenic, VUS, and recurrent somatic variants differ across Pfam domains. Together, these results provide a better characterisation of the most recurrent affected regions in DDRs and could help in the understanding of individual susceptibility to tumour development.

Background: the epidemiological characteristics of acute myocardial infarction have changed in the last decades. Objective: to characterize the patients with acute myocardial infarction admitted to the “Dr. Ernesto Guevara de la Serna” General Teaching Hospital of Las Tunas, from June 2018 through June 2020. Methods: an observational, descriptive and cross-sectional study was carried out at the department of cardiology of the aforementioned hospital and during the period herein stated. The study universe was made up of 754 patients who were admitted with acute coronary syndrome and the study sample consisted of 440 patients with a confirmed diagnosis of acute myocardial infarction. For the data treatment percentage analysis was used for the qualitative variables and arithmetic mean with standard deviation for the quantitative ones. Results: the average age of the patients was 65.9 ± 12.3 years. 64.1% were male and 76.8 % suffered from arterial hypertension. 81.8 % did not have heart failure at admission. 78.4 % presented ST-elevation infarction, thrombolysis was performed on 45.2% and out of them 83.4 % achieved total reperfusion. 31.4% developed left ventricular dysfunction. Hospital mortality was 8.8 %. In more than 90% aspirin, clopidogrel and statins were prescribed at discharge. 20.2% was referred to cardiac rehabilitation. Conclusions: most of the patients were male, with ST-segment elevation myocardial infarction and with a low percentage of reperfusion. Arterial hypertension was the most frequent associated factor. Hospital mortality was low.

Abstract The Chauvet cave (UNESCO World Heritage site, France) is located in the Ardèche Gorge, a unique physical and cultural landscape. Its setting within the gorge—overlooking a meander cutoff containing a natural arch called the Pont d’Arc—is also remarkable. Investigating possible associations between sites’ physical and cultural settings, chronologies of human occupation, and access conditions has become a major theme in archeological research. The present study aims to reconstruct the landscape of the Pont d'Arc meander cutoff during the Upper Paleolithic, when humans were present in the Chauvet Cave. We used uranium-series and electron spin resonance analyses to date the formation of the Pont d’Arc natural arch in the Combe d’Arc meander cutoff, near the Chauvet Cave. Results show that the meander became totally cutoff between 108 and 138 ka (95%). Hence, the natural arch formed before the Upper Paleolithic and the first known human presence in the Chauvet Cave, dated to 37 ka cal BP. These results allowed us to reconstruct a key part of the landscape surrounding the Chauvet Cave when it was being used by Upper-Paleolithic societies.

Abstract Glioblastoma is the most common primary brain tumor. Standard therapy consists of maximum safe resection combined with adjuvant radiochemotherapy followed by chemotherapy with temozolomide, however prognosis is extremely poor. Assessment of the residual tumor after surgery and patient stratification into prognostic groups (i.e., by tumor volume) is currently hindered by the subjective evaluation of residual enhancement in medical images (magnetic resonance imaging [MRI]). Furthermore, objective evidence defining the optimal time to acquire the images is lacking. We analyzed 144 patients with glioblastoma, objectively quantified the enhancing residual tumor through computational image analysis and assessed the correlation with survival. Pathological enhancement thickness on post-surgical MRI correlated with survival (hazard ratio: 1.98, p

Abstract Species diversification in ancient lakes has enabled essential insights into evolutionary theory as they embody an evolutionary microcosm compared to continental terrestrial habitats. We have studied the high-altitude amphipods of the Andes Altiplano using mitogenomic, nuclear ribosomal and single-copy nuclear gene sequences obtained from 36 Hyalella genomic libraries, focusing on species of the Lake Titicaca and other water bodies of the Altiplano northern plateau. Results show that early Miocene South American lineages have recently (late Pliocene or early Pleistocene) diversified in the Andes with a striking morphological convergence among lineages. This pattern is consistent with the ecological opportunities (access to unoccupied resources, initial relaxed selection on ecologically-significant traits and low competition) offered by the lacustrine habitats established after the Andean uplift.

Abstract Schistosomiasis (bilharzia) is a neglected tropical disease caused by parasitic flatworms of the genus Schistosoma, with considerable morbidity in parts of the Middle East, South America, Southeast Asia, in sub-Saharan Africa, and particularly also in Europe. The WHO describes an increasing global health burden with more than 290 million people threatened by the disease and a potential to spread into regions with temperate climates like Corsica, France. The aim of our study was to investigate the influence of S. mansoni infection on colorectal carcinogenic signaling pathways in vivo and in vitro. S. mansoni infection, soluble egg antigens (SEA) and the Interleukin-4-inducing principle from S. mansoni eggs induce Wnt/β-catenin signaling and the protooncogene c-Jun as well as downstream factor Cyclin D1 and markers for DNA-damage, such as Parp1 and γH2a.x in enterocytes. The presence of these characteristic hallmarks of colorectal carcinogenesis was confirmed in colon biopsies from S. mansoni-infected patients demonstrating the clinical relevance of our findings. For the first time it was shown that S. mansoni SEA may be involved in the induction of colorectal carcinoma-associated signaling pathways.

Summary: Background: Visual impairments are a critical medical hurdle to be addressed in modern society. Müller glia (MG) have regenerative potential in the retina in lower vertebrates, but not in mammals. However, in mice, in vivo cell fusion between MG and adult stem cells forms hybrids that can partially regenerate ablated neurons. Methods: We used organotypic cultures of human retina and preparations of dissociated cells to test the hypothesis that cell fusion between human MG and adult stem cells can induce neuronal regeneration in human systems. Moreover, we established a microinjection system for transplanting human retinal organoids to demonstrate hybrid differentiation. Findings: We first found that cell fusion occurs between MG and adult stem cells, in organotypic cultures of human retina as well as in cell cultures. Next, we showed that the resulting hybrids can differentiate and acquire a proto-neural electrophysiology profile when the Wnt/beta-catenin pathway is activated in the adult stem cells prior fusion. Finally, we demonstrated the engraftment and differentiation of these hybrids into human retinal organoids. Interpretation: We show fusion between human MG and adult stem cells, and demonstrate that the resulting hybrid cells can differentiate towards neural fate in human model systems. Our results suggest that cell fusion-mediated therapy is a potential regenerative approach for treating human retinal dystrophies. Funding: This work was supported by La Caixa Health (HR17-00231), Velux Stiftung (976a) and the Ministerio de Ciencia e Innovación, (BFU2017-86760-P) (AEI/FEDER, UE), AGAUR (2017 SGR 689, 2017 SGR 926).

Background: Quality of life is a key outcome after coronary revascularization for ischemic heart disease in the modern era. 6–12 months after coronary revascularization, recovery from the intervention itself, especially coronary artery bypass grafting, is no longer the key determinant of quality of life, while the benefits (or lack thereof) of revascularization become more apparent. Potential determinants of quality of life that are not directly related to the form of intervention have been less frequently examined in this group of patients. Methods: This systematic review examined the association of published contributors to quality of life in patients following coronary revascularization for ischemic heart disease 6–12 months post intervention. Studies that reported such associations were identified and assessed for quality. 11 were included in this review. Quality of life was measured by the Short Forms 36 and 12, and the Seattle Angina Questionnaire. Results: Of potentially modifiable factors, current smoking and hypertension displayed the most consistent associations with reduced quality of life after intervention. Quality of life was positively associated with baseline quality of life and being partnered/married, and negatively associated with female sex, diabetes, and lung or renal disease, and only minimally associated with ejection fraction. Age and prior myocardial infarction displayed conflicting associations. Conclusion: To improve patient outcomes following coronary revascularization, other potentially modifiable non-procedural factors that affect quality of life could be targeted for more intensive management. Smoking cessation and anti-hypertensive therapies treat the most consistent factors associated with reduced quality of life after revascularization.

Objectives: Pakistani women experience higher-than-average rates of certain chronic diseases, including diseases related to sedentary lifestyles. The aim of this study is to explore how first and second-generation Pakistani women living in the Raval, Barcelona, conceive of physical activity, and their barriers and facilitators around participating in physical activity, with the goal of increasing physical activity in this group. Methods: Qualitative research with an intersectional approach. Nine informal interviews with key community informants were conducted from November 2018 to January 2019 to gain background on the topic, using snowball sampling. Eleven individual interviews were conducted from February to June 2019. Seven were with Pakistani women having lived, or currently living, in the neighborhood of the Raval, Barcelona. Four additional interviews were conducted with non-Pakistani women key community informants who have worked extensively with this community. Thematic content analysis was carried out using ATLAS.ti. Results: First-generation Pakistani women generally did not have physical activity present in their daily lives, but by most accounts wished to. Areas that shed light on this included the following: limited economic opportunities and associated living conditions, barriers to social integration, health concepts and access to information, and cultural norms and related gender roles. For the first-generation, gender-related divisions of labor as well as the absence of the concept of self-care were particularly relevant barriers to their participation in physical activity. The experience of immigration-related grief emerged as a transversal theme which overlapped with multiple areas. While both generations expressed a need for the separation of genders during physical activity—as per their cultural interpretation of Islam—the lack of such spaces was highlighted as a principal barrier in physical activity among the second generation. Conclusion: These findings shed light on distinct elements that exert influence in Pakistani women immigrants’ participation in physical activity—among them: social and living conditions, access to public space, and gender-related work distribution and cultural norms—which are in turn influenced by first or second-generation immigration status. Considering the specific needs of both groups when proposing politics and health programs to encourage physical activity is paramount in order to successfully partner with these populations.

Abstract Background After experiencing a sharp growth in COVID-19 cases early in the pandemic, South Korea rapidly controlled transmission while implementing less stringent national social distancing measures than countries in Europe and the USA. This has led to substantial interest in their “test, trace, isolate” strategy. However, it is important to understand the epidemiological peculiarities of South Korea’s outbreak and characterise their response before attempting to emulate these measures elsewhere. Methods We systematically extracted numbers of suspected cases tested, PCR-confirmed cases, deaths, isolated confirmed cases, and numbers of confirmed cases with an identified epidemiological link from publicly available data. We estimated the time-varying reproduction number, R t , using an established Bayesian framework, and reviewed the package of interventions implemented by South Korea using our extracted data, plus published literature and government sources. Results We estimated that after the initial rapid growth in cases, R t dropped below one in early April before increasing to a maximum of 1.94 (95%CrI, 1.64–2.27) in May following outbreaks in Seoul Metropolitan Region. By mid-June, R t was back below one where it remained until the end of our study (July 13th). Despite less stringent “lockdown” measures, strong social distancing measures were implemented in high-incidence areas and studies measured a considerable national decrease in movement in late February. Testing the capacity was swiftly increased, and protocols were in place to isolate suspected and confirmed cases quickly; however, we could not estimate the delay to isolation using our data. Accounting for just 10% of cases, individual case-based contact tracing picked up a relatively minor proportion of total cases, with cluster investigations accounting for 66%. Conclusions Whilst early adoption of testing and contact tracing is likely to be important for South Korea’s successful outbreak control, other factors including regional implementation of strong social distancing measures likely also contributed. The high volume of testing and the low number of deaths suggest that South Korea experienced a small epidemic relative to other countries. Caution is needed in attempting to replicate the South Korean response in populations with larger more geographically widespread epidemics where finding, testing, and isolating cases that are linked to clusters may be more difficult.

An amendment to this paper has been published and can be accessed via the original article.

Alterations in the frequency and intensity of drought events are expected due to climate change and might have consequences for plant metabolism and the development of plant antagonists. In this study, the responses of spring wheat (Triticum aestivum) and one of its major pests, the aphid Sitobion avenae, to different drought regimes were investigated, considering different time points and plant parts. Plants were kept well-watered or subjected to either continuous or pulsed drought. Phloem exudates were collected twice from leaves and once from ears during the growth period and concentrations of amino acids, organic acids and sugars were determined. Population growth and survival of the aphid S. avenae were monitored on these plant parts. Relative concentrations of metabolites in the phloem exudates varied with the time point, the plant part as well as the irrigation regime. Pronounced increases in relative concentrations were found for proline, especially in pulsed drought-stressed plants. Moreover, relative concentrations of sucrose were lower in phloem exudates of ears than in those of leaves. The population growth and survival of aphids were decreased on plants subjected to drought and populations grew twice as large on ears compared to leaves. Our study revealed that changes in irrigation frequency and intensity modulate plant-aphid interactions. These effects may at least partly be mediated by changes in the metabolic composition of the phloem sap.

Abstract Each year, billions of songbirds cross large ecological barriers during their migration. Understanding how they perform this incredible task is crucial to predict how global change may threaten the safety of such journeys. Earlier studies based on radar suggested that most songbirds cross deserts in intermittent flights at high altitude, stopping in the desert during the day, while recent tracking with light loggers suggested diurnal prolongation of nocturnal flights and common non-stop flights for some species. We analyzed light intensity and temperature data obtained from geolocation loggers deployed on 130 individuals of ten migratory songbird species, and show that a large variety of strategies for crossing deserts exists between, but also sometimes within species. Diurnal stopover in the desert is a common strategy in autumn, while most species prolonged some nocturnal flights into the day. Non-stop flights over the desert occurred more frequently in spring than in autumn, and more frequently in foliage gleaners. Temperature recordings suggest that songbirds crossed deserts with flight bouts performed at various altitudes according to species and season, along a gradient ranging from low above ground in autumn to probably >2000 m above ground level, and possibly at higher altitude in spring. High-altitude flights are therefore not the general rule for crossing deserts in migrant songbirds. We conclude that a diversity of migration strategies exists for desert crossing among songbirds, with variations between but also within species.

Chagas disease affects millions of people mainly in Latin America and is a protozoan illness caused by the parasite Trypanosoma cruzi. Chagasic cardiomyopathy is the leading cause of mortality of infected patients, due to compromised electrical and mechanical cardiac function induced by tissue remodeling, especially fibrosis, and lymphocytic infiltration. Some cellular biochemical pathways can be protective to the heart, and we tested if the in vivo activation of the autophagic machinery by rapamycin could reduce parasite-induced myocarditis. Regarding the expression of LC3, an autophagy marker, we observed its upregulation in the cardiac tissue of infected untreated mice. However, after rapamycin treatment, an autophagy inducer, infected mice showed reduced electrical cardiac dysfunctions, myocarditis, cardiac damage, and reduced production of pro-inflammatory cytokines by the heart. On the other hand, the parasite’s life cycle was not affected, and we observed no modulations in cardiac tissue or blood parasitemia. Our data indicate that, at least partially, autophagy induction controls inflammation in the heart¸ illustrating the complexity of the pathways that concur to the development of the infection.

Introduction Malaria infection during pregnancy is an important driver of maternal and neonatal health especially among HIV-infected women. Intermittent preventive treatment in pregnancy (IPTp) with sulphadoxine–pyrimethamine is recommended for malaria prevention in HIV-uninfected women, but it is contraindicated in those HIV-infected on cotrimoxazole prophylaxis (CTXp) due to potential adverse effects. Dihydroartemisinin–piperaquine (DHA–PPQ) has been shown to improve antimalarial protection, constituting a promising IPTp candidate. This trial’s objective is to determine if monthly 3-day IPTp courses of DHA–PPQ added to daily CTXp are safe and superior to CTXp alone in decreasing the proportion of peripheral malaria parasitaemia at the end of pregnancy.Methods and analysis This is a multicentre, two-arm, placebo-controlled, individually randomised trial in HIV-infected pregnant women receiving CTXp and antiretroviral treatment. A total of 664 women will be enrolled at the first antenatal care clinic visit in sites from Gabon and Mozambique. Participants will receive an insecticide-treated net, and they will be administered monthly IPTp with DHA-PPQ or placebo (1:1 ratio) as directly observed therapy from the second trimester of pregnancy. Primary study outcome is the prevalence of maternal parasitaemia at delivery. Secondary outcomes include prevalence of malaria-related maternal and infant outcomes and proportion of adverse perinatal outcomes. Participants will be followed until 6 weeks after the end of pregnancy and their infants until 1 year of age to also evaluate the impact of DHA–PPQ on mother-to-child transmission of HIV. The analysis will be done in the intention to treat and according to protocol cohorts, adjusted by gravidity, country, seasonality and other variables associated with malaria.Ethics and dissemination The protocol was reviewed and approved by the institutional and national ethics committees of Gabon and Mozambique and the Hospital Clinic of Barcelona. Project results will be presented to all stakeholders and published in open-access journals.Trial registration number NCT03671109.