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Abstract Background The architecture and dynamics of T cell populations are critical in orchestrating the immune response to SARS-CoV-2. In our study, we used T Cell Receptor sequencing (TCRseq) to investigate TCR repertoires in 173 post-infection COVID-19 patients. Methods The cohort included 98 mild and 75 severe cases with a median age of 53. We amplified and sequenced the TCR β chain Complementary Determining Region 3 (CDR3b) and performed bioinformatic analyses to assess repertoire diversity, clonality, and V/J allelic usage between age, sex and severity groups. CDR3b amino acid sequence inference was performed by clustering structural motifs and filtering validated reactive CDR3b to COVID-19. Results Our results revealed a pronounced decrease in diversity and an increase in clonal expansion in the TCR repertoires of severe COVID-19 patients younger than 55 years old. These results reflect the observed trends in patients older than 55 years old (both mild and severe). In addition, we identified a significant reduction in the usage of key V alleles (TRBV14, TRBV19, TRBV15 and TRBV6-4) associated with disease severity. Notably, severe patients under 55 years old had allelic patterns that resemble those over 55 years old, accompanied by a skewed frequency of COVID-19-related motifs. Conclusions Present results suggest that severe patients younger than 55 may have a compromised TCR repertoire contributing to a worse disease outcome. Graphical Abstract

Abstract Rifaximin is FDA-approved for treatment of irritable bowel syndrome with diarrhea (IBS-D), but poor solubility may limit its efficacy against microbes in the mucus layer, e.g. Escherichia coli. Here we evaluate adding the mucolytic N-acetylcysteine (NAC) to improve rifaximin efficacy. In a resazurin checkerboard assay, combining rifaximin with NAC had significant synergistic effects in reducing E. coli levels. The optimal rifaximin + NAC combination was then tested in a validated rat model of IBS-D (induced by cytolethal distending toxin [CdtB] inoculation). Rats were inoculated with vehicle and treated with placebo (Control-PBS) or rifaximin + NAC (Control-Rif + NAC, safety), or inoculated with CdtB and treated with placebo (CdtB-PBS), rifaximin (CdtB-Rifaximin), or rifaximin + NAC (CdtB-Rif + NAC) for 10 days. CdtB-inoculated rats (CdtB-PBS) developed wide variability in stool consistency (P = 0.0014) vs. controls (Control-PBS). Stool variability normalized in rats treated with rifaximin + NAC (CdtB-Rif + NAC) but not rifaximin alone (CdtB-Rifaximin). Small bowel bacterial levels were elevated in CdtB-PBS rats but normalized in CdtB-Rif + NAC but not CdtB-Rifaximin rats. E. coli and Desulfovibrio spp levels (each associated with different IBS-D microtypes) were also elevated in CdtB-inoculated (CdtB-PBS) but normalized in CdtB-Rif + NAC rats. Cytokine levels normalized only in CdtB-Rif + NAC rats, in a manner predicted to be associated with reduced diarrhea driven by reduced E. coli. These findings suggest that combining rifaximin with NAC may improve the percentage of IBS-D patients responding to treatment.

Abstract Background The temporal relationships across cardiometabolic diseases (CMDs) were recently conceptualized as the cardiometabolic continuum (CMC), sequence of cardiovascular events that stem from gene-environmental interactions, unhealthy lifestyle influences, and metabolic diseases such as diabetes, and hypertension. While the physiological pathways linking metabolic and cardiovascular diseases have been investigated, the study of the sex and population differences in the CMC have still not been described. Methods We present a machine learning approach to model the CMC and investigate sex and population differences in two distinct cohorts: the UK Biobank (17,700 participants) and the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) (7162 participants). We consider the following CMDs: hypertension (Hyp), diabetes (DM), heart diseases (HD: angina, myocardial infarction, or heart failure), and stroke (STK). For the identification of the CMC patterns, individual trajectories with the time of disease occurrence were clustered using k-means. Based on clinical, sociodemographic, and lifestyle characteristics, we built multiclass random forest classifiers and used the SHAP methodology to evaluate feature importance. Results Five CMC patterns were identified across both sexes and cohorts: EarlyHyp, FirstDM, FirstHD, Healthy, and LateHyp, named according to prevalence and disease occurrence time that depicted around 95%, 78%, 75%, 88% and 99% of individuals, respectively. Within the UK Biobank, more women were classified in the Healthy cluster and more men in all others. In the EarlyHyp and LateHyp clusters, isolated hypertension occurred earlier among women. Smoking habits and education had high importance and clear directionality for both sexes. For ELSA-Brasil, more men were classified in the Healthy cluster and more women in the FirstDM. The diabetes occurrence time when followed by hypertension was lower among women. Education and ethnicity had high importance and clear directionality for women, while for men these features were smoking, alcohol, and coffee consumption. Conclusions There are clear sex differences in the CMC that varied across the UK and Brazilian cohorts. In particular, disadvantages regarding incidence and the time to onset of diseases were more pronounced in Brazil, against woman. The results show the need to strengthen public health policies to prevent and control the time course of CMD, with an emphasis on women.

Abstract Many cancer patients do not benefit from PD-L1/PD-1 blockade immunotherapies. PD-1 and LAG-3 co-upregulation in T-cells is one of the major mechanisms of resistance by establishing a highly dysfunctional state in T-cells. To identify shared features associated to PD-1/LAG-3 dysfunctionality in human cancers and T-cells, multiomic expression profiles were obtained for all TCGA cancers immune infiltrates. A PD-1/LAG-3 dysfunctional signature was found which regulated immune, metabolic, genetic, and epigenetic pathways, but especially a reinforced negative regulation of the TCR signalosome. These results were validated in T-cell lines with constitutively active PD-1, LAG-3 pathways and their combination. A differential analysis of the proteome of PD-1/LAG-3 T-cells showed a specific enrichment in ubiquitin ligases participating in E3 ubiquitination pathways. PD-1/LAG-3 co-blockade inhibited CBL-B expression, while the use of a bispecific drug in clinical development also repressed C-CBL expression, which reverted T-cell dysfunctionality in lung cancer patients resistant to PD-L1/PD-1 blockade. The combination of CBL-B-specific small molecule inhibitors with anti-PD-1/anti-LAG-3 immunotherapies demonstrated notable therapeutic efficacy in models of lung cancer refractory to immunotherapies, overcoming PD-1/LAG-3 mediated resistance.

Abstract In this study, the objective is to understand the causal factors that influence the mental well-being of healthcare workers during the COVID-19 pandemic, using Karasek’s Job Demand Control Social Model (JDCS) as the research framework. For this purpose the database Eurofound’s Living Working and COVID-19 survey is used; it was carried out using sampling design techniques in all 27 countries of the European Union. A sample of 4626 employees in the health sector is used, and structural equation models are specified to analyze causality. The analysis revealed significant associations between several factors: physical and mental work demands (p

The operationalisation of Responsible Research and Innovation is increasingly associated with ethical toolkits. However, scholars remain critical of those toolkits, often referring to them as theoretically problematic, toothless, or too instrumental. Moreover, toolkits imply ideological commitments that are not necessarily made explicit. In this scoping review, we analyse 127 tools designed for technology ethics as part of the RRI Project. We find that (1) these tools tend to frame responsibility as general training or aimed at the development phase of technologies, while monitoring is underrepresented. (2) These toolkits often lack substantive conceptualisations of ethics ignoring contested paradigms. (3) Emerging digital and biotechnologies are over-represented in relation to other socio-technical infrastructures, and (4) there is a risk of a PDF-ization of ethics, as most toolkits are materially constructed as reading material and checklists. We conclude by presenting prompt questions to reflectively reconsider how we design ethical toolkits for technology.

ABSTRACTDiarrhea-predominant irritable bowel syndrome (IBS-D), associated with increased intestinal permeability, inflammation, and small intestinal bacterial overgrowth, can be triggered by acute gastroenteritis. Cytolethal distending toxin B (CdtB) is produced by gastroenteritis-causing pathogens and may underlie IBS-D development, through molecular mimicry with vinculin. Here, we examine the effects of exposure to CdtB alone on gut microbiome composition, host intestinal gene expression, and IBS-D-like phenotypes in a rat model. CdtB-inoculated rats exhibited increased anti-CdtB levels, which correlated with increased stool wet weights, pro-inflammatory cytokines (TNFα, IL2) and predicted microbial metabolic pathways including inflammatory responses, TNF responses, and diarrhea. Three distinct ileal microbiome profiles (microtypes) were identified in CdtB-inoculated rats. The first microtype (most like controls) had altered relative abundance (RA) of genera Bifidobacterium, Lactococcus, and Rothia. The second had lower microbial diversity, higher Escherichia-Shigella RA, higher absolute E. coli abundance, and altered host ileal tissue expression of immune-response and TNF-response genes compared to controls. The third microtype had higher microbial diversity, higher RA of hydrogen sulfide (H2S)-producer Desulfovibrio, and increased expression of H2S-associated pain/serotonin response genes. All CdtB-inoculated rats exhibited decreased ileal expression of cell junction component mRNAs, including vinculin-associated proteins. Significantly, cluster-specific microRNA-mRNA interactions controlling intestinal permeability, visceral hypersensitivity/pain, and gastrointestinal motility genes, including several previously associated with IBS were seen. These findings demonstrate that exposure to CdtB toxin alone results in IBS-like phenotypes including inflammation and diarrhea-like stool, decreased expression of intestinal barrier components, and altered ileal microtypes that influenced changes in microRNA-modulated gene expression and predicted metabolic pathways consistent with specific IBS-D symptoms.

Objectives: The isolated Príncipe is at the malaria pre-elimination stage. Autochthonous clinical cases have been reported sporadically on the island, signaling the possibility of a sizable subpatent (i.e., rapid diagnostic test- and microscopy-negative and polymerase chain reaction [PCR]-positive) parasite reservoir. Methods: Asymptomatic low-density infections were detected by quantitative PCR (qPCR) targeting Plasmodium falciparum multicopy genes (pfr364 and varATS). Positivity rates were assayed for samples surveyed by active case detection (n = 112) and reactive case detection (n = 221) in 2022. Results: qPCR unveiled 70% of low parasitemia carriers, reaching >90% in reactive case detection. The high P. falciparum prevalence was confirmed by the two high-sensitivity qPCR protocols. Higher positivity rates were observed in the localities where most malaria cases were reported in 2022. Most parasitemias were very low (

Abstract Here we studied the entire Atlantic Forest hotspot to investigate whether the effect of different environmental predictors depends on the phylogenetic extension and the biogeographical history of different Atlantic Forest sectors. We used occurrence data of 3,183 plant species with arboreal or arborescent habits. We reconstructed climatic stability across 120,000 years using the Random Forest method. Then, we compared the effect of biogeographical history, topographic, and climatic variables on species richness and phylogenetic diversity using Geographically Weighted Regression (GWR) models. Niche conservatism drives the strength and direction of environmental correlates with tree diversity, interacting with the biogeographical and phylogenetic extension considered. Low current climate seasonalities were the main drivers of species richness and phylogenetic diversity variation across the Atlantic Forest. Whereas in higher phylogenetic extension, topographic heterogeneity increased the number of tree species independent of the sector, deep-past climate stability favored phylogenetic diversity by increasing relict lineages of distant clades in all forests, but with anomalies in the southern sector. This investigation yields substantial evidence that the response of the northern and southern sectors of the Atlantic Forest to identical environmental conditions diverges significantly, providing compelling support for the imprint of phylogenetic heritage in generating non-linear diversity patterns.

Purpose – The growth in the number of nongovernmental organizations (NGOs) worldwide has led to increased competition for donations. A stronger NGO brand equity will make donors more attracted to an organization, compelling them to increase both their donations and their commitment. The goal of this study is to propose a novel donor-based brand equity model. The present study takes into consideration the special characteristics that donors confer to NGOs—specific examples of nonprofit organizations (NPOs) that demand higher moral capital. The suggested framework considers the donor's perspective of NGO brand equity and identifies new dimensions: familiarity (recall, brand strength and brand identification), associations (authenticity, reputation and differentiation) and commitment (attitudinal, emotional) by building on previous NPOs and consumer-based brand equity models. Design/methodology/approach – Based on the analysis of the literature, the authors propose an NGO donor-based brand equity model, which the authors test with a convenience sample of 137 individuals through partial least squares structural equation modeling. Findings – The results of this study demonstrate the positive effects of brand reputation, brand differentiation, brand identification and brand commitment on donor-based brand equity. Practical implications – The novel proposed model will help NGO managers better understand the sources of brand equity from the donor's perspective and more efficiently manage their resources and activities to strengthen their NGO's brand equity. Originality/value – This paper provides a novel, multidimensional NGO donor-based brand equity model that is oriented to the specific characteristics of NGOs; this orientation distinguishes it from previous NPOs and commercial brand equity models. 研究目的 – 隨著全球的非政府組織的數目不斷增加, 爭取捐款的競爭也日趨激烈。一個強大的非政府組織品牌資產, 會吸引捐款者、使其對該組織更為關注, 因而驅使他們增加捐助和支持。本研究擬提出一個新穎的、以捐助者為基礎的品牌資產模型。我們這個建議, 考慮了捐助者賦予非政府組織的一些特徵, 而這些非政府組織是一些強烈要求更高道德資本的特殊例子。我們建議的框架, 考慮了捐助者如何從其角度看待非政府組織的品牌資產, 亦建立了新的層面, 這包括熟悉度 (回憶、品牌強度、品牌識別) 、關聯 (真確性、聲譽、差異化) 、以及支持度 (在態度上的、或在情感上的); 建立這框架, 是透過研究以往的非政府組織、以及以消費者為基礎的品牌資產模型, 並以此為基礎而完成的。 研究設計/方法 – 我們分析有關的文獻, 並以此為基礎, 提出一個以捐助者為基礎的非政府組織的品牌資產模型。我們使用偏最小平方法的結構方程模型, 並測試了隨便抽樣的137個獨立個體, 來試驗這個新模型。 研究結果 – 研究結果顯示, 品牌信譽、品牌差異化、品牌識別和品牌忠誠度, 均會對以捐助者為基礎的品牌資產帶來正面的影響。 實務方面的啟示 – 我們提出的新模型, 讓非政府組織的管理人員能從捐助者的角度、去進一步瞭解品牌資產的來源, 從而更能有效地管理資源和組織的活動, 以便強化組織的品牌資產。 研究的原創性/價值 – 研究提供了以非政府組織特徵為導向的一個、以捐助者為基礎的新穎而俱多層面的非政府組織品牌資產模型。這個研究取向、有別於過往的非營利組織或商業性的品牌資產模型。

Background: Lateral and medial elbow tendinopathies are common soft tissue disorders affecting 1–3% of the general population, causing significant pain and functional impairment in the elbow and upper limb. While often associated with overuse and repetitive strain, their exact etiology, including potential associations with prior injuries in adjacent joints, remains unclear. This preliminary study aims to explore the distribution of lateral and medial elbow tendinopathies and investigate the occurrence of previous lesions in adjacent joints among diagnosed individuals, providing foundational insights for future research. Methods: A multicenter cross-sectional observational study was conducted involving 90 subjects diagnosed with lateral and/or medial elbow tendinopathy. The data collection occurred during the initial consultations, including demographic information, clinical assessments, and history of prior injuries in adjacent joints. Results: Among the sample, 44.4% reported prior injuries to adjacent joints in the affected upper limb, with 45.6% of these injuries identified as musculotendinous in nature. The analysis also showed that the type of elbow tendinopathy was significantly associated with sex (p = 0.01) and occupational origin (p = 0.022). Conclusions: While a notable percentage of the subjects reported prior musculoskeletal injuries in the same limb, the study’s geographic limitations and reliance on self-reported data introduce potential recall bias. These preliminary findings suggest a possible relationship between prior adjacent joint injuries and elbow tendinopathy. Further research with larger sample sizes and more rigorous study design is needed to confirm these observations and explore the underlying mechanisms.

While cytostatic chemotherapy targeting DNA is known to induce genotoxicity, leading to cell cycle arrest and cytokine secretion, the impact of these drugs on fibroblast–epithelial cancer cell communication and metabolism remains understudied. Our research focused on human breast fibroblast RMF-621 exposed to nonlethal concentrations of cisplatin and doxorubicin, revealing reduced proliferation, diminished basal and maximal mitochondrial respirations, heightened mitochondrial ROS and lactate production, and elevated MCT4 protein levels. Interestingly, RMF-621 cells enhanced glucose uptake, promoting lactate export. Breast cancer cells MCF-7 exposed to conditioned media (CM) from drug-treated stromal RMF-621 cells increased MCT1 protein levels, lactate-driven mitochondrial respiration, and a significantly high mitochondrial spare capacity for lactate. These changes occurred alongside altered mitochondrial respiration, mitochondrial membrane potential, and superoxide levels. Furthermore, CM with doxorubicin and cisplatin increased migratory capacity in MCF-7 cells, which was inhibited by MCT1 (BAY-8002), glutamate dehydrogenase (EGCG), mitochondrial pyruvate carrier (UK5099), and complex I (rotenone) inhibitors. A similar behavior was observed in T47-D and ZR-75-1 breast cancer cells. This suggests that CM induces metabolic rewiring involving elevated lactate uptake to sustain mitochondrial bioenergetics during migration. Treatment with the mitochondrial-targeting antioxidant mitoTEMPO in RMF-621 and the addition of an anti-CCL2 antibody in the CM prevented the promigratory MCF-7 phenotype. Similar effects were observed in THP1 monocyte cells, where CM increased monocyte recruitment. We propose that nonlethal concentrations of DNA-damaging drugs induce changes in the cellular environment favoring a promalignant state dependent on mitochondrial bioenergetics.

There is little long-term outcome data on the efficacy of autologous hematopoietic stem cell transplantation (ASCT) in light chain deposition disease (LCDD). We identified 51 LCDD patients in the EBMT registry who had undergone upfront ASCT between 1995 and 2021. The median serum creatinine was 280 μmol/L and 45% required renal replacement therapy (RRT) at time of transplant. The melphalan dose was 100mg/m2 in 23%, 140mg/m2 in 55% and 200 mg/m2 in 21%. The rate of very good partial response or better improved from 41% pre-transplant to 66% at Day +100 post-ASCT. In RRT-independent patients, there was a modest improvement in renal function within the first 3 months; the median eGFR increased from 44 to 51 ml/min/1.73 m2. There was no further change between 3 and 12 months post- ASCT. No patient who was RRT-independent at ASCT became RRT dependent by Day + 100 post-ASCT. Median follow-up post-ASCT was 84 months (IQR: 46-122). At 6-years post ASCT, overall survival (OS) was 88% (95% CI: 78-98%) and PFS was 44% (95% CI: 28-60%). The 2-year cumulative incidence of relapse and non-relapse mortality (NRM) was 17% (95% CI: 6-27%) and 2% (95% CI: 0-6%), respectively. The cumulative incidence of renal transplantation at 4 years after ASCT was 27% (95% CI 13-41) with renal transplantation performed between 6.3 and 52.9 months post-ASCT (median 24.7 months). ASCT represents a feasible option for LCDD patients even if RRT dependent at time of transplant. Outcomes are favourable with low NRM and good long-term OS.

Abstract The main purpose of this study was to evaluate the effect of high‐pressure processing (HPP) and storage temperature on the microbial counts, the instrumental color, and the oxidation stability of sliced dry‐cured Iberian loin from two categories and two leading companies. 600 MPa for 8 min was sufficient to decrease all the microbial counts without affecting the color and the oxidation status, the effect being modulated by the loin category and company, whose effect on those variables was marked. However, the subsequent 90‐day storage softened the initial effect of HPP on microorganisms and allowed a significant effect of HPP to develop on color and oxidation. In addition, the coliform counts were higher after storage at 20°C than at 4°C, suggesting that refrigeration may be needed during long‐term storage to ensure loin safety.

AbstractStarting from the hypothesis that, currently, the potential aesthetic and formal values of engineering are not recognized by society, the main objective of this work was to deepen understanding of the perception of engineering work in Spain and Portugal. For this purpose, a structured survey was developed, and the collected data were subjected to descriptive statistics and non-parametric assessment to evaluate differences among groups. A total of 586 participants were considered in this study, of which 80% had difficulties in recognizing contemporary art, 60% continued to associate it with beauty and 75% acknowledged that they do not understand art or are indifferent to it. The age, education, and urban context of the respondents were factors that influenced levels of knowledge and acceptance of civil engineering work as art, with a clear positioning of architecture as the standard-bearer of its aesthetics. When addressing surveyed interpretation of images, frequency of visits to cultural venues favours the perception of contemporary works of art, while most selected civil works were designed by engineers, which shows that aesthetics and design don’t have to be assigned to other professionals. The vast majority of the participants agree with the notion that engineering enables trust, while about a third of the participants disagree with the idea that art dominates science in engineering work. Finally, there is a broad perception among participants that engineering works can be elevated to a new form of art.

The present study aimed to assess the agreement of three commercially available devices on the measurement of anterior chamber depth (ACD) with and without compensation by central corneal thickness measurement (CCT). Fifty eyes were included in an observational cross-sectional study. Participants underwent a single visit during which devices were used to obtain the inclusion/exclusion (ARK510A, Canon TX-10) and studied (VX-120, Lenstar LS900 and EchoScan US-800) parameters. Based on invasiveness, tests were always performed in the same order by one researcher (to avoid inter-observer variability) and only in the right eye (to avoid overstating the precision of estimates) in each participant. The keratometry, autorefraction, intraocular pressure and anterior chamber angle values were used as inclusion criteria, while the CCT and ACD values were used in the agreement analysis between devices. There was a general and a paired difference in ACD measurements between devices (Greenhouse–Geisser: p ≤ 0.001; Sidak: all p ≤ 0.001). No significant difference was found in ACD measurements compensated by CCT values between the devices (Greenhouse–Geisser: p = 0.200). Pairwise analysis showed a significant difference in VX-120 vs. Lenstar (Sidak: p = 0.021). The differences in ACD measurements compensated by CCT values between the devices were clinically acceptable. Consequently, using these instruments interchangeably in daily routines based on this correction is justified.

Abstract Venous thromboembolism (VTE) is a common, multi-causal disease with potentially serious short- and long-term complications. In clinical practice, there is a need for improved plasma biomarker-based tools for VTE diagnosis and risk prediction. Here we show, using proteomics profiling to screen plasma from patients with suspected acute VTE, and several case-control studies for VTE, how Complement Factor H Related 5 protein (CFHR5), a regulator of the alternative pathway of complement activation, is a VTE-associated plasma biomarker. In plasma, higher CFHR5 levels are associated with increased thrombin generation potential and recombinant CFHR5 enhanced platelet activation in vitro. GWAS analysis of ~52,000 participants identifies six loci associated with CFHR5 plasma levels, but Mendelian randomization do not demonstrate causality between CFHR5 and VTE. Our results indicate an important role for the regulation of the alternative pathway of complement activation in VTE and that CFHR5 represents a potential diagnostic and/or risk predictive plasma biomarker.

Summary: Dorsal and ventral hippocampus serve different functions in cognition and affective behavior, but the underpinnings of this diversity at the cellular and synaptic level are not well understood. We found that the basal level of activin A, a member of the TGF-β family, which regulates hippocampal circuits in a behaviorally relevant fashion, is much higher in dorsal than in ventral hippocampus. Using transgenic mice with a forebrain-specific disruption of activin receptor signaling, we identified the pronounced dorsal-ventral gradient of activin A as a major factor determining the distinct neurophysiologic signatures of dorsal and ventral hippocampus, ranging from pyramidal cell firing, tuning of frequency-dependent synaptic facilitation, to long-term potentiation (LTP), long-term depression (LTD), and de-potentiation. Thus, the strong activin A tone in dorsal hippocampus appears crucial to establish cellular and synaptic phenotypes that are tailored specifically to the respective network operations in dorsal and ventral hippocampus.

Decentralized monitoring methods, which divide the process variables into several blocks and perform local monitoring for each sub-block, have been gaining increasing attention in large-scale plant-wide monitoring due to the complexity of their processes. In such methods, the dynamic nature of the process data is a relevant issue which is not usually managed. Here, a new data-driven distributed dynamic monitoring scheme is proposed to deal with this issue, integrating regression to automatically divide the blocks, a multivariate and dynamic statistical analysis (Canonical Variate Analysis, CVA) to perform local monitoring, and Bayesian inference to achieve the decision making. By constructing sub-blocks using regression, it is possible to identify the most commonly associated variables for every block. Three regression methods are proposed: LASSO (Least Absolute Shrinkage and Selection Operator), which forces the coefficients of the less relevant variables towards zero; Elastic-net, a robust method that is a compromise between Ridge and Lasso regression; and, finally, a non-linear regression method based on the Multilayer Perceptron Network (MLP). Then, the CVA model is implemented for each sub-block to consider the dynamic characteristics of the industrial processes and the Bayesian inference provides a global decision for fault detection. The Tennessee Eastman benchmark validates the efficiency and feasibility of the proposed method regarding some state-of-the-art methods.

Abstract Background The objective of the present study is to describe the characteristics of interstitial pneumonia with autoimmune features (IPAF) patients, to assess the incidence rate of functional respiratory impairment over time and to evaluate the influence of therapeutic alternatives on the prognosis of these patients. Methods A longitudinal observational multicenter study was performed (NEREA registry). It was carried out by a multidisciplinary team in seven Hospitals of Madrid. Patients were included from IPAF diagnosis. Main outcome: poor prognosis as functional respiratory impairment (relative decline in FVC % defined as ≥ 5% every 6 months). Covariates: therapy, sociodemographic, clinical, radiological patterns, laboratory and functional tests. Statistics: Survival techniques were used to estimate IR per 100 patients-semester with their 95% confidence interval [CI]. The influence of covariates in prognosis were analyzed through cox multivariate regression models (hazard ratio (HR) and [CI]). Results 79 IPAF were included, with a mean and a maximum follow-up of 3.17 and 12 years respectively. Along the study, 77.2% received treatment (52 glucocorticoids, 25 mycophenolate, 21 azathioprine, 15 rituximab and 11 antifibrotics). IR was 23.9 [19.9–28.8], and 50% of IPAF developed functional respiratory impairment after 16 months from its diagnosis. Multivariate analysis: usual interstitial pneumonia (UIP) had poorer prognosis compared to non-specific interstitial pneumonia (NSIP) (p = 0.001). In NSIP, positive ANA, increased the risk of poor prognosis. In UIP, glucocorticoids (HR: 0.53 [0.34–0.83]), age (HR: 1.04 [1.01–1.07]), and Ro-antibodies (HR: 0.36 [0.19–0.65]) influenced the prognosis. Conclusions IPAF have functional impairment during the first years of disease. Factors predicting deterioration differ between radiographic patterns. Our real-life study suggests the potential benefit of particular therapies in IPAF.

Purpose: University learning of a group of competencies, also proposed by Bologna, is currently gaining importance: transversal competencies, already included in the OECD Competencies Report (2019) which, common to all university degrees, bring the student closer to a new reality and to a diverse professional future, where sustainable leaders will be one of the fundamental axes of any decision. Design/methodology/approach: The results of a massive survey that was carried out on how university students perceive and value these skills are analyzed, together with a series of qualitative interviews with employers, analyzing what deficiencies exist. Findings: The results show that soft skills are essential for the professional future of students and that they understand and value it. Likewise, it was observed that it is essential to establish a common system of certified recognition of these competences and that universities must develop instruments that facilitate their accreditation in the workplace through digital badges. implications: For its effective development, it is necessary to incorporate in the different degree studies a type of training anchored in the values and attitudes necessary to achieve a fairer and more balanced society, in line with what the Sustainable Development Goals 3, 4 and 8 mark. Originality/value: This study provides a vision of the need for training in certain skills to achieve a rapprochement between the labor market and the university.

The present study aims to use Wood’s model to determine the parameters of individual lactation curves in a local goat population and their factors of variation under a pastoral system. A total of 137,927 records from 432 local goats were collected to assess the impact of litter size, year and month of kidding, herd and the age of the dam on lactation curve parameters. Wood’s model parameters were estimated using non-linear regression, and individual curves were fitted. The characteristics of the lactation curves were computed. The initial yield (A), rate of increase (B) and rate of decline (C) parameters in Wood’s model for local goats were 730 g, 0.26 and 0.09 respectively. The values of peak milk production (PP), peak date (PD) and persistency (PC) were 931.88 g/d, 23.39 days and 91.50%, respectively. Persistence was higher in goats with simple births, while peak production increased by around 0.3 kg for each additional kid. The curve parameters “A” and “C” differed according to the herd and month of kidding (p < 0.05); the age of the dam only had an impact on parameter “A” (p < 0.01). Parameter “B” was not significantly influenced by any of the factors considered (p > 0.05). Correlation coefficients among lactation curve characteristics were ranged from −0.20 to 0.89. Due to a significant negative correlation, selecting for parameter “A” may have an adverse effect on parameter “B”, resulting in a shorter time to reach peak production and less persistency, but an increase in peak production among goats. The curves derived from Wood’s model suggest that the shape of the curve may serve as a basis for herd management planning and to improve local population potentialities.

Rings and gaps are among the most widely observed forms of substructure in protoplanetary disks. A gap–ring pair may be formed when a planet carves a gap in the disk, which produces a local pressure maximum following the gap that traps inwardly drifting dust grains and appears as a bright ring owing to the enhanced dust density. A dust-trapping ring would provide a promising environment for solid growth and possibly planetesimal production via the streaming instability. We present evidence of dust trapping in the bright ring of the planet-hosting disk Elias 2-24, from the analysis of 1.3 and 3 mm Atacama Large Millimeter/submillimeter Array observations at high spatial resolution (0.″029, 4.0 au). We leverage the high spatial resolution to demonstrate that larger grains are more efficiently trapped and place constraints on the local turbulence (8 × 10 ^−4 < α _turb < 0.03) and the gas-to-dust ratio (Σ _g /Σ _d < 30) in the ring. Using a scattering-included marginal probability analysis, we measure a total dust disk mass of ${M}_{\mathrm{dust}}={13.8}_{-0.5}^{+0.7}\times {10}^{-4}\ {M}_{\odot }$ . We also show that at the orbital radius of the proposed perturber the gap is cleared of material down to a flux contrast of 10 ^−3 of the peak flux in the disk.

Some South American countries have ancient traditions that may pose legal problems, such as the consumption of coca leaves, as this can provide positive results for cocaine use after the analysis of biological samples. For this reason, it is necessary to find specific markers that help differentiate legal from illegal consumption, such as tropacocaine, cinnamoylcocaine, and especially hygrine and cuscohygrine. In this work, two techniques for collecting biological samples are compared: the Quantisal® Oral Fluid collection device and passive drooling. Once the samples were collected, they were subjected to solid-phase extraction for subsequent injection into GC-MS. Different validation parameters included in international guides have been studied to evaluate whether the proposed method is valid for the defined purpose, placing special emphasis on the study of the matrix effect and little value on GC-MS analyses. With respect to this parameter, an increase in the signal was found for CUS and t-CIN, but it was not significant for the rest of the substances studied. The recoveries have varied significantly depending on the way of working, being higher when working with standardized areas. After carrying out work with the oral fluid samples collected from laboratory volunteers, the method was applied to two real samples. The results obtained support the need for further research to overcome certain limitations presented by the device.

While the increased availability of traffic data is allowing us to better understand urban mobility, research on data-driven and predictive modeling is also providing new methods for improving traffic management and reducing congestion. In this paper, we present a hybrid predictive modeling architecture, namely GAT-LSTM, by incorporating graph attention (GAT) and long short-term memory (LSTM) networks for handling traffic prediction tasks. In this architecture, GAT networks capture the spatial dependencies of the traffic network, LSTM networks capture the temporal correlations, and the Dayfeature component incorporates time and external information (such as day of the week, extreme weather conditions, holidays, etc.). A key attention block is designed to integrate GAT, LSTM, and the Dayfeature components as well as learn and assign weights to these different components within the architecture. This method of integration is proven effective at improving prediction accuracy, as shown by the experimental results obtained with the PeMS08 open dataset, and the proposed model demonstrates state-of-the-art performance in these experiments. Furthermore, the hybrid model demonstrates adaptability to dynamic traffic conditions, different prediction horizons, and various traffic networks.

Introduction There is a high prevalence of mental health problems among university students. Better prediction and treatment access for this population is needed. In recent years, short-term dynamic factors, which can be assessed using experience sampling methods (ESM), have presented promising results for predicting mental health problems.Methods and analysis Undergraduate students from five public universities in Spain are recruited to participate in two web-based surveys (at baseline and at 12-month follow-up). A subgroup of baseline participants is recruited through quota sampling to participate in a 15-day ESM study. The baseline survey collects information regarding distal risk factors, while the ESM study collects short-term dynamic factors such as affect, company or environment. Risk factors will be identified at an individual and population level using logistic regressions and population attributable risk proportions, respectively. Machine learning techniques will be used to develop predictive models for mental health problems. Dynamic structural equation modelling and multilevel mixed-effects models will be considered to develop a series of explanatory models for the occurrence of mental health problems.Ethics and dissemination The project complies with national and international regulations, including the Declaration of Helsinki and the Code of Ethics, and has been approved by the IRB Parc de Salut Mar (2020/9198/I) and corresponding IRBs of all participating universities. All respondents are given information regarding access mental health services within their university and region. Individuals with positive responses on suicide items receive a specific alert with indications for consulting with a health professional. Participants are asked to provide informed consent separately for the web-based surveys and for the ESM study. Dissemination of results will include peer-reviewed scientific articles and participation in scientific congresses, reports with recommendations for universities’ mental health policy makers, as well as a well-balanced communication strategy to the general public.Study registration osf.io/p7csq.

We present our surgical technique for the reconstruction of the posterolateral corner of the knee. It is a tibia- and fibular-based reconstruction technique. Most of these procedures require the use of 2 tendons (autograft or allograft). In our technique, a single semitendinosus tendon is required, making the procedure more suitable if the surgeon prefers the use of autograft or when there is no access to a tissue bank. This is even more important in the setting of multiligament knee injuries. The most defining feature of this modification is the possibility of achieving the desired graft tension in a progressive and independent way, due to the use of 3 adjustable-loop cortical suspension devices.

This research aims to investigate PAI teachers' strategies through learning designs at Muhammadiyah Middle Schools. This type of research is descriptive qualitative field research. Muhammadiyah Middle School in Yogyakarta is the site of this research. Three PAI teachers served as research subjects, with the research objects being PAI teacher strategies, student learning behavior, and Islamic education learning designs. Techniques for gathering data include observation, interviews, and documentation. Techniques for data analysis include data reduction, data presentation, and conclusion. According to the study's findings, there are two learning design strategies: planning and implementing learning. The planning design is adjusted to content standards and is passed down in the syllabus to the Learning Implementation Plan (RPP). At the same time, implementation is divided into three steps: opening, core activities, and closing. It begins with asking students how they are and brainstorming material. The core activities include communicating learning objectives, delivering material, and assessing students' attitudes, knowledge, and skills. The final activity, closing, is filled out by the teacher with learning reflections and conclusions.

Abstract Background Entrepreneurial intention is considered to be the best predictor of entrepreneurial behaviour. The Theory of Planned Behaviour (TPB) explains the degree of correlation between variables such as entrepreneurial intention, perceived feasibility and perceived desirability. Knowing the entrepreneurial intention of students of Health Sciences will help to guide and promote effective university policies to support entrepreneurship. The authors aimed to analyse the entrepreneurial intention of university students in the field of Health Sciences. Methods A cross-sectional study was conducted in the Faculties of Health Sciences of two public universities of Southern Spain. 1518 students of different degrees of Health Sciences (Physiotherapy, Podiatry, Dentistry, Nursing and Occupational Therapy), from first to fourth year. An online structured questionnaire was used, the Entrepreneurial Event Model (EEM) adapted to the Spanish context. This measurement model was completed with 8 items from the Motivated Strategies for Learning Questionnaire-MSLQ. Results The hypothetical model showed that perceived desirability and perceived feasibility were positive and significant predictors of entrepreneurial intention. Perceived desirability showed an indirect effect on entrepreneurial intention through perceived feasibility. Expectation of success and self-efficacy had no direct effect on entrepreneurial intention. Conclusions Perceived desirability and perceived feasibility are related to entrepreneurial intention in Health Sciences students.

BackgroundEstimating and analyzing trends and patterns of health loss are essential to promote efficient resource allocation and improve Peru’s healthcare system performance.MethodsUsing estimates from the Global Burden of Disease (GBD), Injuries, and Risk Factors Study (2019), we assessed mortality and disability in Peru from 1990 to 2019. We report demographic and epidemiologic trends in terms of population, life expectancy at birth (LE), mortality, incidence, prevalence, years of life lost (YLLs), years lived with disability (YLDs), and disability-adjusted life-years (DALYs) caused by the major diseases and risk factors in Peru. Finally, we compared Peru with 16 countries in the Latin American (LA) region.ResultsThe Peruvian population reached 33.9 million inhabitants (49.9% women) in 2019. From 1990 to 2019, LE at birth increased from 69.2 (95% uncertainty interval 67.8–70.3) to 80.3 (77.2–83.2) years. This increase was driven by the decline in under-5 mortality (−80.7%) and mortality from infectious diseases in older age groups (+60 years old). The number of DALYs in 1990 was 9.2 million (8.5–10.1) and reached 7.5 million (6.1–9.0) in 2019. The proportion of DALYs due to non-communicable diseases (NCDs) increased from 38.2% in 1990 to 67.9% in 2019. The all-ages and age-standardized DALYs rates and YLLs rates decreased, but YLDs rates remained constant. In 2019, the leading causes of DALYs were neonatal disorders, lower respiratory infections (LRIs), ischemic heart disease, road injuries, and low back pain. The leading risk factors associated with DALYs in 2019 were undernutrition, high body mass index, high fasting plasma glucose, and air pollution. Before the COVID-19 pandemic, Peru experienced one of the highest LRIs-DALYs rates in the LA region.ConclusionIn the last three decades, Peru experienced significant improvements in LE and child survival and an increase in the burden of NCDs and associated disability. The Peruvian healthcare system must be redesigned to respond to this epidemiological transition. The new design should aim to reduce premature deaths and maintain healthy longevity, focusing on effective coverage and treatment of NCDs and reducing and managing the related disability.

ABSTRACT BACKGROUND: Gallbladder polyps are becoming a common finding in ultrasound. The management has to consider the potential risk of malignant lesions. AIMS: The aim of this study was to analyze the ultrasound findings in patients undergoing cholecystectomy due to gallbladder polyps and compare them for histopathological findings (HPs). METHODS: Patients with an ultrasonographic diagnosis of gallbladder polyp and who underwent cholecystectomy from 2007 to 2020 were included in the study. RESULTS: A total of 447 patients were included, of whom 58% were women. The mean age was 45±12 years. The mean size of polyps in US was 7.9±3.6 mm. Notably, 9% of polyps were greater than 10 mm, and single polyps were significantly larger than the multiple ones (p=0.003). Histopathological findings confirmed the presence of polyps in 88.4%, with a mean size of 4.8±3.4 mm. In all, 16 cases were neoplastic polyps (4.1%), 4 of them being malignancies, and all were single and larger than 10 mm. We found a significant correlation between ultrasound and histopathological findings polyp size determination (r=0.44; p

Summary: Background: Gallbladder cancer (GBC) is rare but aggressive. The extent of surgical intervention for different GBC stages is non-uniform, ranging from cholecystectomy alone to extended resections including major hepatectomy, resection of adjacent organs and routine extrahepatic bile duct resection (EBDR). Robust evidence here is lacking, however, and survival benefit poorly defined. This study assesses factors associated with recurrence-free survival (RFS), overall survival (OS) and morbidity and mortality following GBC surgery in high income countries (HIC) and low and middle income countries (LMIC). Methods: The multicentre, retrospective Operative Management of Gallbladder Cancer (OMEGA) cohort study included all patients who underwent GBC resection across 133 centres between 1st January 2010 and 31st December 2020. Regression analyses assessed factors associated with OS, RFS and morbidity. Findings: On multivariable analysis of all 3676 patients, wedge resection and segment IVb/V resection failed to improve RFS (HR 1.04 [0.84–1.29], p = 0.711 and HR 1.18 [0.95–1.46], p = 0.13 respectively) or OS (HR 0.96 [0.79–1.17], p = 0.67 and HR 1.48 [1.16–1.88], p = 0.49 respectively), while major hepatectomy was associated with worse RFS (HR 1.33 [1.02–1.74], p = 0.037) and OS (HR 1.26 [1.03–1.53], p = 0.022). Furthermore, EBDR (OR 2.86 [2.3–3.52], p

IntroductionLegionnaires’ Disease is a pneumonia caused by Legionella spp., currently treated empirically with fluoroquinolones and macrolides. In this study, we aim to describe the antibiotic susceptibility pattern of environmental Legionella recovered in the south of Portugal.MethodsMinimal inhibitory concentration (MIC) determination of 57 Legionella isolates (10 Lp sg 1, 32, Lp sg 2-14 15 L. spp) was achieved by broth microdilution, as described by EUCAST, for azithromycin, clarithromycin, ciprofloxacin, levofloxacin, and doxycycline. ResultsFluoroquinolones were the most active antibiotic, displaying the lowest MIC values in contrast to doxycycline which had the highest. MIC90 and epidemiological cut-off (ECOFF) values were, respectively, 0.5/1 mg/L for azithromycin, 0.125/0.25 mg/L for clarithromycin, 0.064/0.125 mg/L for ciprofloxacin, 0.125/0.125 mg/L for levofloxacin and 16/32 mg/L for doxycycline.DiscussionMIC distributions were higher than reported by EUCAST for all antibiotics. Interestingly, two phenotypically resistant isolates with high-level quinolone resistance were identified. This is the first time that MIC distributions, lpeAB and tet56 genes have been investigated in Portuguese environmental isolates of Legionella.

A single congenital muscular torticollis (CMT) is a postural musculoskeletal deformity and is characterized by the shortening or stiffness of the sternocleidomastoid muscle. The reported incidence of CMT ranges from 0.2% to 2%. The objective is to evaluate the effect of physical therapy programs on CMT. For the search, PubMed, Scopus, Web of Science, PEDro and Cochrane databases were used. Randomized controlled trials published between 2018 and 2023 have been included. This study follows the PRISMA 2020 statement and has been registered in the PROSPERO database. Finally, six studies were included. The cervical range of motion (ROM) in rotation was the most analyzed variable, followed by the ultrasound evaluation; one of the studies included the analysis of children’s motor development with the Alberta scale. All research found benefits associated with soft tissue mobilization, passive stretching techniques and manual therapy of the cervical spine. In conclusion, it is possible to recommend manual therapy and passive stretching techniques for the treatment of CMT, with significant results on the cervical ROM.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive neoplasm with very poor patient survival outcomes despite available treatments. There is an urgent need for new potential treatment options and novel biomarkers for these patients. Delta-like canonical Notch ligand 3 (DLL3) interacts with the Notch receptor and causes inhibition of Notch signaling, which confers a survival advantage to PDAC cells. Thus, DLL3 expression could affect cell survival, and its inhibition could increase a patient’s survival. To test this hypothesis, a survival analysis was conducted using the progression-free and overall survival from two independent datasets of PDAC patients, with one using mRNA z-score levels and the other using the Hscore protein expression level; both were carried out using a log-rank test and plotted using Kaplan–Meier curves. DLL3 at the mRNA expression level showed an association between high mRNA expression and both a longer progression-free survival (PFS) and overall survival (OS) of patients. Then, we designed a retrospective study with resected PDAC samples. Our primary objective with this dataset was to assess the relationship between PFS and OS and DLL3 protein expression. The secondary assessment was to provide a rationale for the use of anti-DLL3-based treatments in combination with immunotherapy that is supported by the link between DLL3 and other factors that are involved in immune checkpoints. The survival analyses revealed a protective effect of high DLL3 protein expression levels in both PFS and OS. Interestingly, high DLL3 protein expression levels were significantly correlated with PD-L1/2 and negatively correlated with NOTCH1. Therefore, DLL3 could be considered a biomarker for better prognosis in resectable PDAC patients as well as a therapeutic biomarker for immunotherapy response. These facts set a rationale for testing anti-DLL3-based treatments either alone or combined with immunotherapy or other NOTCH1 inhibitors.

Abstract The International Staging System (ISS) and the Revised International Staging System (R-ISS) are commonly used prognostic scores in multiple myeloma (MM). These methods have significant gaps, particularly among intermediate-risk groups. The aim of this study was to improve risk stratification in newly diagnosed MM patients using data from three different trials developed by the Spanish Myeloma Group. For this, we applied an unsupervised machine learning clusterization technique on a set of clinical, biochemical and cytogenetic variables, and we identified two novel clusters of patients with significantly different survival. The prognostic precision of this clusterization was superior to those of ISS and R-ISS scores, and appeared to be particularly useful to improve risk stratification among R-ISS 2 patients. Additionally, patients assigned to the low-risk cluster in the GEM05 over 65 years trial had a significant survival benefit when treated with VMP as compared with VTD. In conclusion, we describe a simple prognostic model for newly diagnosed MM whose predictions are independent of the ISS and R-ISS scores. Notably, the model is particularly useful in order to re-classify R-ISS score 2 patients in 2 different prognostic subgroups. The combination of ISS, R-ISS and unsupervised machine learning clusterization brings a promising approximation to improve MM risk stratification.

Higher education students are usually not familiar with active learning, especially students from degrees not related to education. This is not the case of primary and secondary education students, that have been involved in several active methodologies during their studies. This paper presents a game-based learning proposal that was implemented during a cryptography outreach session with high school students and with third-year students from computer science undergraduate degree. Both groups were separately involved in the activity, and they found the solution of all riddles and problems. Students behavior was similar, as all of them were engage with the game, where they learnt cryptography, while they were engaged in the active learning session acquiring transversal and specific skills.

Introduction and Objectives: Acute-on-chronic liver failure (ACLF) is a severe clinical entity with organ failures and high short-term mortality. To Date, few ACLF reports have been published in Latin America. This study aimed to characterize patients with ACLF, identifying triggers, organ failure, and survival at 30, 90, and 180 days compared to patients with decompensated cirrhosis without ACLF. Materials and Methods: Retrospective study of decompensated cirrhotic patients hospitalized (between 2017-2019) in three centers in Chile. We evaluated transplant-free survival using Kaplan-Meier curves and Cox-regression. Results: 398 patients were included, a median age of 65.3±11.7-year-old, 50.5% female, 91 (22.9%) presented ACLF (8% ACLF-1, 6.3% ACLF-2, 8.6% ACLF-3); 6.6% underwent liver transplantation. ACLF patients were younger (63.6 vs. 66.0 years; p=0.045), had higher MELD-Na scores (27 [23-32] vs. 17 [13-23]; p

BackgroundApproximately 13.8% and 6.1% of coronavirus disease 2019 (COVID-19) patients require hospitalization and sometimes intensive care unit (ICU) admission, respectively. There is no biomarker to predict which of these patients will develop an aggressive stage that we could improve their quality of life and healthcare management. Our main goal is to include new markers for the classification of COVID-19 patients.MethodsTwo tubes of peripheral blood were collected from a total of 66 (n = 34 mild and n = 32 severe) samples (mean age 52 years). Cytometry analysis was performed using a 15-parameter panel included in the Maxpar® Human Monocyte/Macrophage Phenotyping Panel Kit. Cytometry by time-of-flight mass spectrometry (CyTOF) panel was performed in combination with genetic analysis using TaqMan® probes for ACE2 (rs2285666), MX1 (rs469390), and TMPRSS2 (rs2070788) variants. GemStone™ and OMIQ software were used for cytometry analysis.ResultsThe frequency of CD163+/CD206- population of transitional monocytes (T-Mo) was decreased in the mild group compared to that of the severe one, while T-Mo CD163-/CD206- were increased in the mild group compared to that of the severe one. In addition, we also found differences in CD11b expression in CD14dim monocytes in the severe group, with decreased levels in the female group (p = 0.0412). When comparing mild and severe disease, we also found that CD45- [p = 0.014; odds ratio (OR) = 0.286, 95% CI 0.104–0.787] and CD14dim/CD33+ (p = 0.014; OR = 0.286, 95% CI 0.104–0.787) monocytes were the best options as biomarkers to discriminate between these patient groups. CD33 was also indicated as a good biomarker for patient stratification by the analysis of GemStone™ software. Among genetic markers, we found that G carriers of TMPRSS2 (rs2070788) have an increased risk (p = 0.02; OR = 3.37, 95% CI 1.18–9.60) of severe COVID-19 compared to those with A/A genotype. This strength is further increased when combined with CD45-, T-Mo CD163+/CD206-, and C14dim/CD33+.ConclusionsHere, we report the interesting role of TMPRSS2, CD45-, CD163/CD206, and CD33 in COVID-19 aggressiveness. This strength is reinforced for aggressiveness biomarkers when TMPRSS2 and CD45-, TMPRSS2 and CD163/CD206, and TMPRSS2 and CD14dim/CD33+ are combined.

Substitution of fish-based ingredients may alter the nutritional profile of the feeds, including the vitamin contents, ultimately leading to unbalanced vitamin supply. Vitamin A plays an essential role in epithelium preservation, cell differentiation, reproduction, and vision. It also intervenes in skeletogenesis through chondrocytes development. Therefore, low levels of vitamin A may cause poor growth and abnormal bone development among other symptoms. Besides, in gilthead seabream excess vitamin A altered bone structure and homeostasis, indicating that an upper level for vitamin A in feeds for this species must be defined. For this purpose, a practical plant-based diet (FM 10% and FO 6%) containing five increasing levels of vitamin A (24,000, 26,000, 27,000, 31,000, and 37,000 IU/kg) supplemented as retinyl acetate was formulated to identify the effects of high levels of vitamin A for gilthead seabream juveniles. The trial was conducted with 450 total fish distributed into 15 tanks, where each diet was tested in triplicates for 70 days. At the end of the trial, samples were taken for analyses of vitamin A—relevant markers. At the end of the trial the high levels of vitamin A supplementation did not cause a reduction in growth, whereas no significant effect was observed for the feed efficiency, specific growth rate, and feed convertion ratio. Although not significant, retinol content in liver showed a tendency to increase with the elevation of dietary vitamin A levels. Although minor, the highest level of vitamin A dietary content (37,000 IU/kg) caused a significant increase in caudal vertebrae partial fusion as well as caudal vertebrae malformations. Increasing dietary vitamin A was related to a reduction in the occurrence of microhemorrhages in the liver and a reduction in the presence of eosinophils associated to the pancreas. Overall, the results of the present study suggested that gilthead seabream juveniles fed a plant-based diet are able to tolerate very high levels of vitamin A supplementation when supplemented as retinyl acetate. Nevertheless, further supplementation should be avoided in order to reduce the prevalence of anomalies affecting the caudal vertebrae.

The aggressiveness of melanoma and lack of effective therapies incite the discovery of novel strategies. Recently, a new dual acting hybrid molecule (HM), combining a triazene and a ʟ-tyrosine analogue, was synthesized. HM was designed to specifically be activated by tyrosinase, the enzyme involved in melanin biosynthesis and overexpressed in melanoma. HM displayed remarkable superior antiproliferative activity towards various cancer cell lines compared with temozolomide (TMZ), a triazene drug in clinical use, that acts through DNA alkylation. In B16-F10 cells, HM induced a cell cycle arrest at phase G0/G1 with a 2.8-fold decrease in cell proliferation index. Also, compared to control cells, HM led to a concentration-dependent reduction in tyrosinase activity and increase in caspase 3/7 activity. To maximize the therapeutic performance of HM in vivo, its incorporation in long blood circulating liposomes, containing poly(ethylene glycol) (PEG) at their surface, was performed for passively targeting tumour sites. HM liposomes (LIP HM) exhibited high stability in biological fluids. Preclinical studies demonstrated its safety for systemic administration and in a subcutaneous murine melanoma model, significantly reduced tumour progression. In a metastatic murine melanoma model, a superior antitumour effect was also observed for mice receiving LIP HM, with markedly reduction of lung metastases compared to positive control group (TMZ). Biodistribution studies using 111In-labelled LIP HM demonstrated its ability for passively targeting tumour sites, thus correlating with the high therapeutic effect observed in the two experimental murine melanoma models. Overall, our proposed nanotherapeutic strategy was validated as an effective and safe alternative against melanoma.