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Wilson disease (WD) is a genetic disorder of copper metabolism caused by mutations in the ATP7B gene resulting in toxic copper accumulation in several organs. WD can manifest as liver disease, a progressive neurological disorder, a psychiatric illness, or a combination of these. Other clinical manifestations can also occur. Diagnosis is challenging and typically requires a range of biochemical tests, imaging, genetic testing for ATP7B, and/or liver biopsy. WD is treatable with chelating agents, such as d-penicillamine and trientine, and/or zinc salts alongside with dietary copper restriction. Liver transplantation may be indicated in WD patients with severe hepatic disease, and cautiously considered in patients with neurological WD. Treatment success highly depends on patient adherence and treatment persistence. Therefore, effective interventions for improving patient adherence and close monitoring are key for preventing WD progression. In Portugal, there are no reference centers for WD, and patients are dispersed across numerous medical specialists. This review aimed to summarize the most recent and relevant information for the diagnosis, treatment, and monitoring of WD in Portugal, as well as possible interventions for stimulating adherence to treatment.

Abstract Angiogenesis, vital for tumor growth and metastasis, is a promising target in cancer therapy. Natural compounds offer potential as antiangiogenic agents with reduced toxicity. This review provides a comprehensive overview of natural product‐based antiangiogenic therapies, focusing on molecular mechanisms and therapeutic potential. A systematic search identified relevant articles from 2019 to 2023. Various natural compounds, including polyphenols, terpenes, alkaloids, cannabinoids, omega‐3 fatty acids, polysaccharides, proteins, and carotenoids, were investigated for their antiangiogenic properties. Challenges such as dose standardization, routes of administration, and potential side effects remain. Further studies, including in‐depth animal models and human epidemiological studies, must elucidate clinical efficacy and safety. Synergistic effects with current antiangiogenic therapies, such as bevacizumab and tyrosine kinase inhibitors, should be explored. Additionally, the potential hormone‐dependent effects of compounds like genistein highlight the need for safety evaluation. In conclusion, natural products hold promise as adjunctive therapies to conventional antineoplastic drugs in modulating angiogenesis in cancer. However, robust clinical trials are needed to validate preclinical findings and ensure safety and efficacy.

Aim: Aloysia citrodora has a long history of traditional use in treating various ailments. This study evaluated the in vivo chemopreventive efficacy and systemic toxicity of an extract of A. citrodora in a transgenic mouse model of HPV16 (human papillomavirus type 16)-induced cancer. Methods: The experiment involved six groups (n = 5): group 1 (G1, wild-type (WT), water), group 2 (G2, HPV, water), group 3 (G3, WT, 0.013 g/mL), group 4 (G4, HPV, 0.006 g/mL), group 5 (G5, HPV, 0.008 g/mL), and group 6 (G6, HPV, 0.013 g/mL). Throughout the assay, humane endpoints, body weight, food, and water consumption were recorded weekly. The internal organs and skin of the mice were collected for analysis after they were sacrificed. Toxicological parameters that were studied included hematological and biochemical blood markers, splenic and hepatic histology, and hepatic oxidative stress. Results: A. citrodora extract seems to reduce the incidence of dysplastic and in situ carcinoma skin lesions induced by HPV16 in this model, suggesting that dietary supplementation with concentrations of 0.008 g/mL and 0.013 g/mL may have beneficial chemopreventive effects. Conclusions: The extract did not induce any concentration-dependent toxicological effects on any of the parameters included in the study, indicating a favorable toxicological profile under these experimental conditions.

Abstract The exchange of trace gases between the atmosphere and the ocean plays a key role in the Earth’s climate. Fluxes at the air-sea interface are affected mainly by wind blowing over the ocean and seawater temperature and salinity changes. This study aimed to quantify the use of CO $$_{2}$$ 2 partial pressure (pCO $$_{2}$$ 2 ) measurements at different depths (1, 5, and 10 m) in ocean surface layers to determine CO $$_{2}$$ 2 fluxes (FCO $$_{2}$$ 2 ) and to investigate the impacts of wind-sheltered and wind-exposed regions on the carbon budget. Vertical profiles of temperature, salinity, and pCO $$_{2}$$ 2 were considered during a daily cycle. pCO $$_{2}$$ 2 profiles exhibited relatively high values during sunny hours, associated with relatively high sea temperatures. However, the largest FCO $$_{2}$$ 2 corresponded with higher wind speeds. Estimated fluxes between measurements at 1 and 10 m depths decreased by 71% in the sheltered region and 44% in the exposed region. According to the SOCAT dataset, at a depth of 5 m, the Atlantic basin emits approximately 0.29 Tg month $$^{-1}$$ - 1 of CO $$_{2}$$ 2 to the atmosphere; nevertheless, our estimates suggest that FCO $$_{2}$$ 2 at the surface is 12.02 Tg month $$^{-1}$$ - 1 , which is 97.6% greater than that at 5 m depth. Therefore, future studies should consider sampling depth to adequately estimate the FCO $$_{2}$$ 2 .

Abstract The lack of representative in vitro models recapitulating human tendon (patho)physiology is among the major factors hindering consistent progress in the knowledge‐based development of adequate therapies for tendinopathy.Here, an organotypic 3D tendon‐on‐chip model is designed that allows studying the spatiotemporal dynamics of its cellular and molecular mechanisms.Combining the synergistic effects of a bioactive hydrogel matrix with the biophysical cues of magnetic microfibers directly aligned on the microfluidic chip, it is possible to recreate the anisotropic architecture, cell patterns, and phenotype of tendon intrinsic (core) compartment. When incorporated with vascular‐like vessels emulating the interface between its intrinsic‐extrinsic compartments, crosstalk with endothelial cells are found to drive stromal tenocytes toward a reparative profile. This platform is further used to study adaptive immune cell responses at the onset of tissue inflammation, focusing on interactions between tendon compartment tenocytes and circulating T cells.The proinflammatory signature resulting from this intra/inter‐cellular communication induces the recruitment of T cells into the inflamed core compartment and confirms the involvement of this cellular crosstalk in positive feedback loops leading to the amplification of tendon inflammation.Overall, the developed 3D tendon‐on‐chip provides a powerful new tool enabling mechanistic studies on the pathogenesis of tendinopathy as well as for assessing new therapies.

Abstract Recent studies have shown that the microbiome can impact cancer development, progression, and response to therapies suggesting microbiome-based approaches for cancer characterization. As cancer-related signatures are complex and implicate many taxa, their discovery often requires Machine Learning approaches. This review discusses Machine Learning methods for cancer characterization from microbiome data. It focuses on the implications of choices undertaken during sample collection, feature selection and pre-processing. It also discusses ML model selection, guiding how to choose an ML model, and model validation. Finally, it enumerates current limitations and how these may be surpassed. Proposed methods, often based on Random Forests, show promising results, however insufficient for widespread clinical usage. Studies often report conflicting results mainly due to ML models with poor generalizability. We expect that evaluating models with expanded, hold-out datasets, removing technical artifacts, exploring representations of the microbiome other than taxonomical profiles, leveraging advances in deep learning, and developing ML models better adapted to the characteristics of microbiome data will improve the performance and generalizability of models and enable their usage in the clinic.

This study explores a nanoemulsion formulated with açaí seed oil, known for its rich fatty acid composition and diverse biological activities. This study aimed to characterise a nanoemulsion formulated with açaí seed oil and explore its cytotoxic effects on HeLa and SiHa cervical cancer cell lines, alongside assessing its antioxidant and toxicity properties both in vitro and in vivo. Extracted from fruits sourced in Brazil, the oil underwent thorough chemical characterization using gas chromatography–mass spectrometry. The resulting nanoemulsion was prepared and evaluated for stability, particle size, and antioxidant properties. The nanoemulsion exhibited translucency, fluidity, and stability post centrifugation and temperature tests, with a droplet size of 238.37, PDI -9.59, pH 7, and turbidity 0.267. In vitro assessments on cervical cancer cell lines revealed antitumour effects, including inhibition of cell proliferation, migration, and colony formation. Toxicity tests conducted in cell cultures and female Swiss mice demonstrated no adverse effects of both açaí seed oil and nanoemulsion. Overall, açaí seed oil, particularly when formulated into a nanoemulsion, presents potential for cancer treatment due to its bioactive properties and safety profile.

Immunotherapy has emerged as a promising approach for cancer treatment, with oncolytic adenoviruses showing power as immunotherapeutic agents. In this study, we investigated the immunotherapeutic potential of an adenovirus construct expressing CXCL9, CXCL10, or IL-15 in clear cell renal cell carcinoma (ccRCC) tumor models. Our results demonstrated robust cytokine secretion upon viral treatment, suggesting effective transgene expression. Subsequent analysis using resistance-based transwell migration and microfluidic chip assays demonstrated increased T-cell migration in response to chemokine secretion by infected cells in both 2D and 3D cell models. Flow cytometry analysis revealed CXCR3 receptor expression across T-cell subsets, with the highest percentage found on CD8+ T-cells, underscoring their key role in immune cell migration. Alongside T-cells, we also detected NK-cells in the tumors of immunocompromised mice treated with cytokine-encoding adenoviruses. Furthermore, we identified potential immunogenic antigens that may enhance the efficacy and specificity of our armed oncolytic adenoviruses in ccRCC. Overall, our findings using ccRCC cell line, in vivo humanized mice, physiologically relevant PDCs in 2D and patient-derived organoids (PDOs) in 3D suggest that chemokine-armed adenoviruses hold promise for enhancing T-cell migration and improving immunotherapy outcomes in ccRCC. Our study contributes to the development of more effective ccRCC treatment strategies by elucidating immune cell infiltration and activation mechanisms within the tumor microenvironment (TME) and highlights the usefulness of PDOs for predicting clinical relevance and validating novel immunotherapeutic approaches. Overall, our research offers insights into the rational design and optimization of viral-based immunotherapies for ccRCC.

ABSTRACTThe high background of host RNA poses a major challenge to metatranscriptome analysis of human samples. Hence, metatranscriptomics has been mainly applied to microbe-rich samples, while its application in human tissues with low ratio of microbial to host cells has yet to be explored. Since there is no computational workflow specifically designed for the taxonomic and functional analysis of this type of samples, we propose an effective metatranscriptomics strategy to accurately characterize the microbiome in human tissues with a low ratio of microbial to host content. We experimentally generated synthetic samples with well-characterized bacterial and host cell compositions, and mimicking human samples with high and low microbial loads. These synthetic samples were used for optimizing and establishing the workflow in a controlled setting. Our results show that the integration of the taxonomic analysis of optimized Kraken 2/Bracken with the functional analysis of HUMAnN 3 in samples with low microbial content, enables the accurate identification of a large number of microbial species with a low false-positive rate, while improving the detection of microbial functions. The effectiveness of our metatranscriptomics workflow was demonstrated in synthetic samples, simulated datasets, and most importantly, human gastric tissue specimens, thus providing a proof of concept for its applicability on mucosal tissues of the gastrointestinal tract. The use of an accurate and reliable metatranscriptomics approach for human tissues with low microbial content will expand our understanding of the functional activity of the mucosal microbiome, uncovering critical interactions between the microbiome and the host in health and disease.

Abstract Background Germline mutations of E-cadherin contribute to hereditary diffuse gastric cancer (HDGC) and congenital malformations, such as oral facial clefts (OFC). However, the molecular mechanisms through which E-cadherin loss-of-function triggers distinct clinical outcomes remain unknown. We postulate that E-cadherin-mediated disorders result from abnormal interactions with the extracellular matrix and consequent aberrant intracellular signalling, affecting the coordination of cell migration. Methods Herein, we developed in vivo and in vitro models of E-cadherin mutants associated with either OFC or HDGC. Using a Drosophila approach, we addressed the impact of the different variants in cell morphology and migration ability. By combining gap closure migration assays and time-lapse microscopy, we further investigated the migration pattern of cells expressing OFC or HDGC variants. The adhesion profile of the variants was evaluated using high-throughput ECM arrays, whereas RNA sequencing technology was explored for identification of genes involved in aberrant cell motility. Results We have demonstrated that cells expressing OFC variants exhibit an excessive motility performance and irregular leading edges, which prevent the coordinated movement of the epithelial monolayer. Importantly, we found that OFC variants promote cell adhesion to a wider variety of extracellular matrices than HDGC variants, suggesting higher plasticity in response to different microenvironments. We unveiled a distinct transcriptomic profile in the OFC setting and pinpointed REG1A as a putative regulator of this outcome. Consistent with this, specific RNAi-mediated inhibition of REG1A shifted the migration pattern of OFC expressing cells, leading to slower wound closure with coordinated leading edges. Conclusions We provide evidence that E-cadherin variants associated with OFC activate aberrant signalling pathways that support dynamic rearrangements of cells towards improved adaptability to the microenvironment. This proficiency results in abnormal tissue shaping and movement, possibly underlying the development of orofacial malformations. Graphical Abstract

Most patients with muscle‐invasive bladder cancer (MIBC) are not cured with platinum chemotherapy. Up‐regulation of nuclear factor kappa light‐chain enhancer of activated B cells (NF‐κB) is a major mechanism underlying chemoresistance, suggesting that its pharmacological inhibition may increase platinum efficacy. NF‐κB signaling was investigated in two patient cohorts. The Cancer Genome Atlas (TCGA) was used to correlate NF‐κB signaling and patient survival. The efficacy of cisplatin plus the NF‐κB inhibitor dimethylaminoparthenolide (DMAPT) versus cisplatin or DMAPT alone was tested in vitro. Xenografted and immunocompetent MIBC mouse models were studied in vivo. Platinum‐naive claudin‐low MIBC showed constitutive NF‐κB signaling and this was associated with reduced disease‐specific survival in TCGA patients. Chemotherapy up‐regulated NF‐κB signaling and chemoresistance‐associated genes, including SPHK1, PLAUR, and SERPINE1. In mice, DMAPT significantly improved the efficacy of cisplatin in both models. The combination preserved body weight, renal function, and morphology, reduced muscle fatigue and IL‐6 serum levels, and did not aggravate immuno‐hematological toxicity compared with cisplatin alone. These data provide a rationale for combining NF‐κB inhibition with platinum‐based chemotherapy and conducting a clinical trial in MIBC patients.

Two monoterpenoid lactones, loliolide (1) and epi-loliolide (2), were isolated from the crude dichloromethane extract of a microalga, Thalassiosira sp.). The structures of loliolide (1) and epi-loliolide (2) were elucidated by 1D and 2D NMR analysis, as well as a comparison of their 1H or/and 13C NMR data with those reported in the literature. In the case of loliolide (1), the absolute configurations of its stereogenic carbons were confirmed by X-ray analysis, whereas those of epi-loliolide (2) were determined by NOESY correlations. Loliolide (1) and epi-loliolide (2) were tested for their growth inhibitory activity against two Gram-positive (Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212) and two Gram-negative (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) bacteria, as well as one clinical isolate (E. coli SA/2, an extended-spectrum β-lactamase producer-ESBL) and two environmental isolates, S. aureus 74/24, a methicillin-resistant (MRSA), and E. faecalis B3/101, a vancomycin-resistant (VRE) isolates. The results showed that none of the tested compounds exhibited antibacterial activity at the highest concentrations tested (325 μM), and both revealed low antioxidant activity, with ORAC values of 2.786 ± 0.070 and 2.520 ± 0.319 µmol TE/100 mg for loliolide (1) and epi-loliolide (2), respectively.

Additive manufacturing has a strong potential to produce sound metal–polymer joints using controlled polymer deposition on a metallic substrate. In this way, this study aimed to explore the morphological and mechanical properties of metal–polymer joints produced through material-extrusion-based AM using a pin-based macro-mechanical interlocking mechanism. Joints were fabricated with polylactic acid deposited onto a heated aluminium alloy substrate to form the connection. The optimisation process was focused on improving the printing parameters and pin geometries to reduce voids and enhance joint integrity. The results indicate that optimised samples exhibit superior mechanical resistance, achieving a maximum load improvement with an overall strength increase of 368.97% compared to non-optimised joints. A combined pin geometry (50% cylindrical, 50% conical) was found to be the most effective. Morphological analysis confirmed uniform polymer deposition, ensuring reliable joint performance. These findings underscore the critical role of geometric optimisation in enhancing the strength and durability of metal–polymer joints in AM applications.

Fucus spp. seaweeds thrive in the cold temperate waters of the northern hemisphere, specifically in the littoral and sublittoral regions along rocky shorelines. Moreover, they are known to be a rich source of bioactive compounds. This study explored the valorization of Fucus spiralis through the extraction of bioactives and polysaccharides (PSs) for food applications and biostimulant use. The bioactives were extracted using microwave hydrodiffusion and gravity (MHG), where the condition of 300 W for 20 min resulted in the highest total phenolic content and antioxidant activity of the extract. Cellular assays confirmed that the extract, at 0.5 mg/mL, was non-cytotoxic to HaCat cells. Polysaccharides (PSs) were extracted from the remaining biomass. The residue from this second extraction contained 1.5% protein and 13.35% carbohydrates. Additionally, the free amino acids and minerals profiles of both solid residues were determined. An edible film was formulated using alginate (2%), PS-rich Fucus spiralis extract (0.5%), and F. spiralis bioactive-rich extract (0.25%). The film demonstrated significant antioxidant properties, with ABTS and DPPH values of 221.460 ± 10.389 and 186.889 ± 36.062 µM TE/mg film, respectively. It also exhibited notable physical characteristics, including high water vapor permeability (11.15 ± 1.55 g.mm.m−2.day−1.kPa−1) and 100% water solubility. The residues from both extractions of Fucus spiralis exhibited biostimulant (BS) effects on seed germination and seedling growth. BSs with PSs enhanced pea germination by 48%, while BSs without PSs increased the root dry weight of rice and tomato by 53% and up to 176%, respectively, as well as the shoot dry weight by up to 38% and up to 74%, respectively. These findings underscore the potential of Fucus spiralis within the framework of a circular economy, wherein both extracted bioactives and post-extraction by-products can be used for sustainable agriculture and food applications.

Summary: Corticospinal neurons (CSNs) synapse directly on spinal neurons, a diverse assortment of cells with unique structural and functional properties necessary for body movements. CSNs modulating forelimb behavior fractionate into caudal forelimb area (CFA) and rostral forelimb area (RFA) motor cortical populations. Despite their prominence, the full diversity of spinal neurons targeted by CFA and RFA CSNs is uncharted. Here, we use anatomical and RNA sequencing methods to show that CSNs synapse onto a remarkably selective group of spinal cell types, favoring inhibitory populations that regulate motoneuron activity and gate sensory feedback. CFA and RFA CSNs target similar spinal neuron types, with notable exceptions that suggest that these populations differ in how they influence behavior. Finally, axon collaterals of CFA and RFA CSNs target similar brain regions yet receive highly divergent inputs. These results detail the rules of CSN connectivity throughout the brain and spinal cord for two regions critical for forelimb behavior.

Summary: The brain can generate actions, such as reaching to a target, using different movement strategies. We investigate how such strategies are learned in a task where perched head-fixed mice learn to reach to an invisible target area from a set start position using a joystick. This can be achieved by learning to move in a specific direction or to a specific endpoint location. As mice learn to reach the target, they refine their variable joystick trajectories into controlled reaches, which depend on the sensorimotor cortex. We show that individual mice learned strategies biased to either direction- or endpoint-based movements. This endpoint/direction bias correlates with spatial directional variability with which the workspace was explored during training. Model-free reinforcement learning agents can generate both strategies with similar correlation between variability during training and learning bias. These results provide evidence that reinforcement of individual exploratory behavior during training biases the reaching strategies that mice learn.

The impact of 15% dietary inclusion of Spirulina (Arthrospira platensis) in broiler chickens was explored, focusing on blood cellular components, systemic metabolites and hepatic lipid and mineral composition. From days 14 to 35 of age, 120 broiler chickens were divided and allocated into four dietary treatments: a standard corn and soybean meal-based diet (control), a 15% Spirulina diet, a 15% extruded Spirulina diet, and a 15% Spirulina diet super-dosed with an enzyme blend (0.20% porcine pancreatin plus 0.01% lysozyme). The haematological analysis revealed no significant deviations (p > 0.05) in blood cell counts across treatments, suggesting that high Spirulina inclusion maintains haematological balance. The systemic metabolic assessment indicated an enhanced antioxidant capacity in birds on Spirulina diets (p

Abstract Malignant pleural mesothelioma (MPM) is an aggressive tumor with a poor prognosis. As the available therapeutic options show a lack of efficacy, novel therapeutic strategies are urgently needed. Given its T-cell infiltration, we hypothesized that MPM is a suitable target for therapeutic cancer vaccination. To date, research on mesothelioma has focused on the identification of molecular signatures to better classify and characterize the disease, and little is known about therapeutic targets that engage cytotoxic (CD8+) T cells. In this study we investigate the immunopeptidomic antigen-presented landscape of MPM in both murine (AB12 cell line) and human cell lines (H28, MSTO-211H, H2452, and JL1), as well as in patients’ primary tumors. Applying state-of-the-art immuno-affinity purification methodologies, we identify MHC I-restricted peptides presented on the surface of malignant cells. We characterize in vitro the immunogenicity profile of the eluted peptides using T cells from human healthy donors and cancer patients. Furthermore, we use the most promising peptides to formulate an oncolytic virus-based precision immunotherapy (PeptiCRAd) and test its efficacy in a mouse model of mesothelioma in female mice. Overall, we demonstrate that the use of immunopeptidomic analysis in combination with oncolytic immunotherapy represents a feasible and effective strategy to tackle untreatable tumors.

Background: Teratomas are rare types of germ cell neoplasms composed of various differentiated or undifferentiated tissues. Case Description: A 25-week-old female control FVB/n mouse in a 4-week toxicity study presented abdominal distension and poor body condition. It was euthanized, and the necropsy exam revealed a large mass connected to the tip of the right uterine horn, occupying the entire abdominal cavity. Microscopically, this mass showed areas of epidermal differentiation, with laminated keratin and sebaceous glands, differentiation into respiratory and digestive epithelium, cartilage, bone, and extensive areas of differentiation into the nervous tissue, being classified as an ovarian teratoma. Conclusion: As far as authors know, the occurrence of ovarian teratomas in FVB/n mouse strain has never been previously described. [Open Vet J 2023; 13(9.000): 1223-1227]

This technical note aims to characterize and contextualize the creation of a new collection of identified skeletons stored in the University Institute of Health Sciences—CESPU. This project presents unique and innovative pedagogical opportunities, with students actively involved in the cleaning and processing of the bones and antemortem/postmortem information gathering, as well as in anthropology research projects. This XXI Century Identified Skeletal Collection is composed of 98 complete adult skeletons of both sexes and is still expanding, being the 10th contemporary collection available in Portugal. The skeletons consist exclusively of Portuguese nationals who died between 1946 and 2007, and the 42% that have been processed so far are in good preservation. Comparative learning outcomes after implementing this active learning strategy are not yet available. Improvements are expected since more participation and enthusiasm in classes and research were perceived.

The advancement of engineering and the emergence of increasingly individualized and specific applications have heightened the need to develop advanced manufacturing processes capable of delivering all of the required characteristics [...]

Controlling root movement is one of the greatest challenges in orthodontic treatment with aligners, like Invisalign® aligners. Cone Beam Computed Tomography (CBCT) integration into ClinCheck®, enabling bone and root visualisation, allows a more accurate follow-up of the teeth position. This study aims to compare torque measurements of the upper central incisors with and without CBCT and relate them to the upper incisor inclination and facial biotype. In a sample of 70 teeth, torque measurements were obtained by importing images into AutoCAD® software (version 2024). The angle between the tooth’s long axis with CBCT duplicate and the tooth’s long axis without CBCT was obtained to assess the difference. Statistically significant differences between torque measurements with and without CBCT were found, as well as between these measurements and the inclination of the upper incisors. No statistically significant differences were found among the facial biotypes. The average values of 27.8° ± 3.4° and 21.5° ± 3.2° were obtained for the angle between the axes. Torque without CBCT was lower than torque with CBCT, for the same tooth. The angle between the axes had a similar mean for both teeth. CBCT integration into ClinCheck® allows for a more correct torque measurement.

The joining of AA6082-T6 and polyamide 6 using pinless friction stir spot welding was investigated in this study. The influence of the clamping frame geometry was studied and the welds produced were characterized based on their morphology and mechanical performance. The morphological analysis was evaluated based on the comparison of the different joining areas and on the presence of defects in the resolidified layer of the polymer. In turn, the mechanical performance of the joints was evaluated by tensile-shear testing. Additionally, the influence of plunge depth parameter was studied for the clamping frame geometry providing the best mechanical performance. While the clamping frame geometry had a greater impact on the size of the joining areas, therefore influencing the micro-mechanical interlocking mechanisms, the plunge depth mainly affected the plunging of the aluminium into the polymeric material, therefore affecting the macro-mechanical interlocking mechanism. The strongest joints, which failed for a load of about 2700 N, were produced with the clamping frame geometry that restricted the welding zone the least, and used the highest plunge depth.

The digital era is opening countless possibilities in “Sport Sciences”; “Public, Environmental, and Occupational Health”; and “Physical Therapy, Sports Therapy, and Rehabilitation” areas [...]

Within the framework of sustainable supply chain management and logistics, this work tackles the complex challenge of optimizing inventory levels across varied storage facilities. It introduces a comprehensive many-objective optimization model designed to minimize holding costs, energy consumption, and shortage risk concurrently, thereby integrating sustainability considerations into inventory management. The model incorporates the distinct energy consumption profiles associated with various storage types and evaluates the influence of stock levels on energy usage. Through an examination of a 60-day production schedule, the dynamic relationship between inventory levels and operational objectives is investigated, revealing a well-defined set of optimal solutions that highlight the trade-off between energy savings and shortage risk. Employing a 30-day rolling forward analysis with daily optimization provides insights into the evolving nature of inventory optimization. Additionally, the model is extended to encompass a five-objective optimization by decomposing shortage risk, offering a nuanced comprehension of inventory risks. The outcomes of this research provide a range of optimal solutions, empowering supply chain managers to make informed decisions that strike a balance among cost, energy efficiency, and supply chain resilience.

Abstract Emerging evidence of the relationship between the microbiome composition and the development of numerous diseases, including cancer, has led to an increasing interest in the study of the human microbiome. Technological breakthroughs regarding DNA sequencing methods propelled microbiome studies with a large number of samples, which called for the necessity of more sophisticated data-analytical tools to analyze this complex relationship. The aim of this work was to develop a machine learning-based approach to distinguish the type of cancer based on the analysis of the tissue-specific microbial information, assessing the human microbiome as valuable predictive information for cancer identification. For this purpose, Random Forest algorithms were trained for the classification of five types of cancer—head and neck, esophageal, stomach, colon, and rectum cancers—with samples provided by The Cancer Microbiome Atlas database. One versus all and multi-class classification studies were conducted to evaluate the discriminative capability of the microbial data across increasing levels of cancer site specificity, with results showing a progressive rise in difficulty for accurate sample classification. Random Forest models achieved promising performances when predicting head and neck, stomach, and colon cancer cases, with the latter returning accuracy scores above 90% across the different studies conducted. However, there was also an increased difficulty when discriminating esophageal and rectum cancers, failing to differentiate with adequate results rectum from colon cancer cases, and esophageal from head and neck and stomach cancers. These results point to the fact that anatomically adjacent cancers can be more complex to identify due to microbial similarities. Despite the limitations, microbiome data analysis using machine learning may advance novel strategies to improve cancer detection and prevention, and decrease disease burden.

Neurons coordinate their activity to produce an astonishing variety of motor behaviors. Our present understanding of motor control has grown rapidly thanks to new methods for recording and analyzing populations of many individual neurons over time. In contrast, current methods for recording the nervous system’s actual motor output – the activation of muscle fibers by motor neurons – typically cannot detect the individual electrical events produced by muscle fibers during natural behaviors and scale poorly across species and muscle groups. Here we present a novel class of electrode devices (‘Myomatrix arrays’) that record muscle activity at unprecedented resolution across muscles and behaviors. High-density, flexible electrode arrays allow for stable recordings from the muscle fibers activated by a single motor neuron, called a ‘motor unit,’ during natural behaviors in many species, including mice, rats, primates, songbirds, frogs, and insects. This technology therefore allows the nervous system’s motor output to be monitored in unprecedented detail during complex behaviors across species and muscle morphologies. We anticipate that this technology will allow rapid advances in understanding the neural control of behavior and identifying pathologies of the motor system.

Abstract Ulva lactuca is a seaweed with antinutritional cell wall for monogastrics. Carbohydrate-Active enZymes (CAZymes) supplementation can potentially cause its disruption. This study evaluates four diets: Ctrl—control diet; UL—control + 7% U. lactuca (wild caught, powdered form); ULR—UL + 0.005% Rovabio® Excel AP; ULU—UL + 0.01% ulvan lyase on piglets’ haematologic and serologic profiles, hepatic lipids and minerals. White blood cells and lymphocytes reached the highest values in piglets fed UL compared to control, and to control and ULR; respectively (P 0.05). While hepatic n-3 PUFA increased in piglets fed with seaweed diets due to increment of beneficial 22:5n-3 and 22:6n-3 fatty acids (P 0.05). ULR reduced α-tocopherol levels (P = 0.036) and increased serum potassium levels (P

Objective: This study examines the asymmetric impact of both internal (military, education, and health expenditures) and external (trade opening and foreign direct investment) factors that contribute to poverty reduction. Methodology: To find an asymmetric relationship between the proposed variables, we used a non-linear ARDL co-integration approach for the period ranging from 1981-2019. Findings: The findings of the study confirm the asymmetric impact of internal (education, military, health expenditures, quality of governance) and external (foreign direct investment, openness) factors on poverty. The finding confirms that ignoring nonlinear or asymmetric properties of macroeconomic variables may mislead inferences. This study has policy implications for government officials to reduce poverty. Novelty: theeconomic theory of poverty is studied from different perspectives by using internal and external factors that have direct and indirect effects on poverty. Furthermore, for in-depth analysis, a nonlinear approach is used to determine which factor has a strong contribution to eliminating poverty. Doi: 10.28991/ESJ-2023-07-03-07 Full Text: PDF

Abstract Seaweeds, such as Laminaria digitata, are a sustainable alternative to conventional feedstuffs for weaned piglet diets, improving their health and mitigating environmental impacts. L. digitata has a complex cell wall that can be difficult for monogastrics to digest. However, carbohydrate-active enzymes (CAZymes) such as Rovabio® Excel AP and alginate lyase can help break down these polysaccharides and render intracellular nutrients more accessible. This study aimed to evaluate the impact of 10% L. digitata feed inclusion and CAZyme supplementation on piglet blood cells, serum metabolites, liver lipid and mineral profiles. Forty weaned piglets were randomly assigned to one of four diets (n = 10 each): a control diet, 10% L. digitata (LA), 10% L. digitata + 0.005% Rovabio® Excel AP (LAR), and 10% L. digitata + 0.01% alginate lyase (LAL). After two weeks of trial, animals were slaughtered and liver and blood serum samples taken for analysis. The results showed that the LA and LAL diets increased blood lymphocytes, IgG and IgM, and decreased serum lipids, improving both cellular and humoral immune response and cardiovascular health. Dietary CAZymes reversed the anti-inflammatory and hematopoietic effects. Additionally, cortisol levels were reduced with seaweed inclusion compared to the control diet (P

Forensic sciences aggregate a series of scientific and technological domains that aid the judiciary and judicial system. Despite the highly specialized and qualified professionals taking a role in forensic practice, this paradigmatic integration of law, science, and technology often leads them to face significant ethical challenges. Indeed, forensic sciences hold a unique position due to the social and legal implications inherent to forensic expertise. The potential interference with personal rights and freedoms requires the professional practice of the forensic specialist to be guided by alignment with scientific-technical competence under the seal of the highest ethical principles. However, the absence of a specific statutory regulation blurs the performance of the forensic professional and may harm the professional credibility but also the scientific domain as a whole. The existence of ethical pillars is essential to support professional practice, following international recommendations in this regard. Herein, we propose a framework for a code of ethics and conduct that is based on the professional particularities characterizing the forensic practice but also considers the ethical issues that are mandatory to ensure high levels of reliability and credibility of forensic specialists.

Lemon is a fruit rich in antioxidant properties and has several health benefits, namely the reduction of skin edema and anticarcinogenic properties, which are due to its high content of bioactive compounds. Melatonin can improve and preserve the properties of lemon for longer and also has health benefits. The aim of this study was to evaluate the effects of oral administration of lemon juice after melatonin treatment on murinometric parameters of wild-type (WT) mice and transgenic mice carrying human papillomavirus (HPV). Two trials were performed for oral administration of the lemon extract compound: in drinking water and in diet. First of all, lemons were treated by immersion with melatonin at 10 mM. Then, lemons were squeezed, and the juice obtained was freeze-dried and stored to be subsequently added to drinking water or diet, according to the assay. Thus, mice were divided into eight groups in the drink assay (each with n = 5): group 1 (G1, WT, control), group 2 (G2, WT, 1 mL lemon), group 3 (G3, WT, 1.5 mL lemon), group 4 (G4, WT, 2 mL lemon), group 5 (G5, HPV16, control), group 6 (G6, HPV16, 1 mL lemon) group 7 (G6, HPV16, 1.5 mL lemon) and group 8 (G6, HPV16, 2 mL lemon). The diet assay was divided into four groups: group 1 (G1, WT, control), group 2 (G2, WT, 4 mL lemon), group 3 (G3, HPV16, control) and group 4 (G4, HPV16, 4 mL lemon). In the drink assay, the highest concentration of melatonin (308 ng/100 mL) was for groups 4 and 8, while in the food assay, there was only one concentration of melatonin (9.96 ng/g) for groups 2 and 4. Both trials lasted 30 days. During this time, body weight, food and water were recorded. Afterward, they were sacrificed, and samples were collected for different analyses. At the concentrations used, the lemon juice with melatonin had no adverse effects on the animals’ health and showed a positive outcome in modifying weight gain and enhancing antioxidant activity in mice. Moreover, a reduction in the incidence of histological lesions was observed in treated animals. Further research is needed to better understand the effects of lemon extract on health and treatment outcomes in this animal model.

The use of by-products as a source of bioactive compounds with economic added value is one of the objectives of a circular economy. The olive oil industry is a source of olive pomace as a by-product. The olive pomace used in the present study was the exhausted olive pomace, which is the by-product generated from the air drying and subsequent hexane extraction of residual oil from the olive pomace. The objective was to extract bioactive compounds remaining in this by-product. Various types of green extraction were used in the present study: solvent extraction (water and hydroalcoholic); ultrasound-assisted extraction; Ultra-Turrax-assisted extraction; and enzyme-assisted extraction (cellulase; viscoenzyme). The phenolic profile of each extract was determined using HPLC-DAD and the total phenolic content (TPC) and antioxidant activity (ABTS, DPPH, and ORAC) were determined as well. The results showed significant differences in the yield of extraction among the different methods used, with the enzyme-assisted, with or without ultrasound, extraction presenting the highest values. The ultrasound-assisted hydroethanolic extraction (USAHE) was the method that resulted in the highest content of the identified phenolic compounds: 2.021 ± 0.29 mg hydroxytyrosol/100 mg extract, 0.987 ± 0.09 mg tyrosol/100 mg extract, and 0.121 ± 0.005 mg catechol/100 mg extract. The conventional extraction with water at 50 °C produced the best results for TPC and antioxidant activity of the extracts. The extracts from the USAHE were able to inhibit Gram-positive bacteria, especially Bacillus cereus, showing 67.2% inhibition at 3% extract concentration.

The development of sustainable materials from the valorization of waste is a good alternative to reducing the negative environmental impact of plastic packaging. The objectives of this study were to develop and characterize pectin-based composite films incorporated with cork or cork with either coffee grounds or walnut shells, as well as to test the films’ genotoxicity, antioxidant properties, and biodegradation capacity in soil and seawater. The addition of cork, coffee grounds, or walnut shells modified the films’ characteristics. The results showed that those films were thicker (0.487 ± 0.014 mm to 0.572 ± 0.014 mm), more opaque (around 100%), darker (L* = 25.30 ± 0.78 to 33.93 ± 0.84), and had a higher total phenolic content (3.17 ± 0.01 mg GA/g to 4.24 ± 0.02 mg GA/g). On the other hand, the films incorporated only with cork showed higher values of elongation at break (32.24 ± 1.88% to 36.30 ± 3.25%) but lower tensile strength (0.91 ± 0.19 MPa to 1.09 ± 0.08 MPa). All the films presented more heterogeneous and rougher microstructures than the pectin film. This study also revealed that the developed films do not contain DNA-reactive substances and that they are biodegradable in soil and seawater. These positive properties could subsequently make the developed films an interesting eco-friendly food packaging solution that contributes to the valorization of organic waste and by-products, thus promoting the circular economy and reducing the environmental impact of plastic materials.

We present novel velocimetry algorithms based on the hybridization of correlation-based Particle Image Velocimetry (PIV) and a combination of Lucas–Kanade and Liu–Shen optical flow (OpF) methods. An efficient Aparapi/OpenCL implementation of those methods is also provided in the accompanying open-source QuickLabPIV-ng tool enabled with a Graphical User Interface (GUI). Two different options of hybridization were developed and tested: OpF as a last step, after correlation-based PIV, and OpF as a substitute for sub-pixel interpolation. Hybridization increases the spatial resolution of PIV, enabling the characterization of small turbulent scales and the computation of key turbulence parameters such as the rate of dissipation of turbulent kinetic energy. The method was evaluated using both synthetic and real databases, representing flows that exhibit a variety of locally isotropic homogeneous turbulent scales. The proposed hybrid PIV-OpF results in a 3-fold increase in the PIV density for synthetic images. The analysis of power spectral density functions and auto-correlation demonstrated the impact of PIV image quality on the accuracy of the method and its ability to extend the turbulence range. We discuss the challenges posed by optical noise and tracer density in the quality of the vector map density.

Purpose: This article aims to present successful practices in the management of training processes based on virtual reality and augmented reality, namely a strategy for evaluating the process with the principle of continuous improvement in mind, and monitoring its performance in terms of productivity and waste levels. It is proposed to apply Statistical Process Control (SPC) tools to develop control charts for monitoring individual events (i-charts). Design/methodology/approach: The methodology is based on a case study developed in an industrial project and is guided by a literature review on Work-Based Learning (WBL) and SPC. Findings: The developed work shows that SPC tools are suitable for supporting decision making in situations where the data to be analyzed is generated by human-computer interactions, e.g., involving students and virtual learning environments. Originality/value: The innovative aspect presented in the article lies in the evaluation of the effectiveness of pedagogical resources arranged in simulation environments based on virtual and augmented reality. The accumulated knowledge about the application of SPC in service areas, and others that demand data analysis, reinforces the hypothesis of the suitability of its application in the case presented. This is an original application of SPC, normally used in business processes quality control, but which in this case is applied in an innovative way to the evaluation of industrial training processes, with the same spirit for which it was designed, i.e. to provide the means to manage the quality of a process.

Abstract Kinship analysis from very low-coverage ancient sequences has been possible up to the second degree with large uncertainties. We propose a new, accurate, and fast method, correctKin, to estimate the kinship coefficient and the confidence interval using low-coverage ancient data. We perform simulations and also validate correctKin on experimental modern and ancient data with widely different genome coverages (0.12×–11.9×) using samples with known family relations and known/unknown population structure. Based on our results, correctKin allows for the reliable identification of relatedness up to the 4th degree from variable/low-coverage ancient or badly degraded forensic whole genome sequencing data.

Abstract Objectives The present study aims to characterize the spinal balance (SB) in young adults with Schmorl nodes (SN). Methods A cross-sectional study was conducted on a sample of 47 young adults. Lumbar magnetic resonance imaging (MRI) was used to divide the patients into an SN group and a control group. Standing full spine radiographs were used to compare the spinopelvic SB parameters between groups: sagittal vertical axis, thoracic kyphosis, lumbar lordosis (LL), pelvic incidence (PI), pelvic tilt (PT), and sacral slope (SS). Results The LL and SS values were significantly lower in patients with SN when compared with the control group (54.5° versus 64.3°; 36.2° versus 41.4°, respectively). No significant differences were observed for the other parameters. Significant correlations were found in both groups between LL and SS; PI and PT; and PI and SS. Conclusions Young adults with SN have associated SB modifications, particularly lower LL and SS values, when compared with a control group. This flatter profile resembles that observed in patients with lower back pain and early disc pathology. We believe that SNs are relevant clinical findings that should prompt the study of the SB of a patient, as it may uncover variations associated with early disc degeneration. Level of Evidence III

Abstract In this study, we analysed the impact of dietary inclusion of Chlorella vulgaris and carbohydrases on general health, redox status, immune response, liver lipids and metabolites in weaned piglets. Forty-four male piglets were allocated into four diets: control (n = 11), CH (control diet with 5% CH, n = 10), CH+R (control diet with 5% CH plus 0.005% Rovabio Excel AP, n = 10), and CH+M (control diet with 5% CH plus 0.01% of a pre-selected four-CAZyme mixture, n = 11). After 15 days of trial, animals were slaughtered and samples of blood and liver collected. Spectrophotometry methods and commercial kits were used to determine blood parameters and gas and liquid chromatography for hepatic fatty acid and chlorophylls profiles, respectively. While total, LDL- and VLDL-cholesterol were increased by CH, the opposite was recorded for HDL-cholesterol (p

ABSTRACT Objective: The risk of malignancy and diagnostic accuracy of fine-needle aspiration biopsy (FNAB) of thyroid nodules (TN) with diameters ≥ 3-4 cm remains controversial. However, some groups have indicated surgical treatment in these patients regardless of the FNAB results. We aimed to evaluate the diagnostic accuracy of the FNAB in systematically resected ≥4 cm TN and if the risk of malignancy is higher in these patients. Subjects and methods: We retrospectively evaluated 138 patients (142 nodules) with TN with diameters ≥4 cm who underwent thyroidectomy. Results: The FNAB results were nondiagnostic/unsatisfactory (ND/UNS) in 2.1% of the cases and benign in 51.4%. They indicated atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS) in 23.9% of cases, follicular neoplasia/suspicious for a follicular neoplasm (FN/SFN) in 9.2%, suspicion of malignancy (SUS) in 8.5%, and malignant in 4.9%. The histopathological analysis after thyroidectomy revealed a thyroid cancer rate of 100% in the FNABs classified as malignant, 33.3% in SUS cases, 7.7% in FN/SFN, 17.6% in AUS/FLUS, and 4.1% in benign FNABs. None of the ND/UNS FNABs were malignant. The global malignancy diagnosis was 14.8% (n = 21). However, the rate of false negatives for FNAB was low (4.1%). Conclusion: We showed that the risk of malignancy in nodules with diameters ≥4 cm was higher compared to the risk of thyroid cancer in TN in general. However, we found a low rate of false-negative cytological results; therefore, our data do not justify the orientation of routine resection for these larger nodules.

Crithmum maritimum L. (sea fennel), an edible xerophyte of coastal habitats, is considered an emerging cash crop for biosaline agriculture due to its salt-tolerance ability and potential applications in the agri-food sector. Here, the nutritional value and bioactive properties of sea fennel are described. Sea fennel leaves, flowers, and schizocarps are composed of carbohydrates (>65%) followed by ash, proteins, and lipids. Sea fennel’s salty, succulent leaves are a source of omega-6 and omega-3 polyunsaturated fatty acids, especially linoleic acid. Extracts obtained from flowers and fruits/schizocarps are rich in antioxidants and polyphenols and show antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermis, Candida albicans, and Candida parapsilosis. Plant material is particularly rich in sodium (Na) but also in other nutritionally relevant minerals, such as calcium (Ca), chlorine (Cl), potassium (K), phosphorus (P), and sulfur (S), beyond presenting a potential prebiotic effect on Lactobacillus bulgaricus and being nontoxic to human intestinal epithelial Caco-2 model cells, up to 1.0% (w/v). Hence, the rational use of sea fennel can bring nutrients, aroma, and flavor to culinary dishes while balancing microbiomes and contributing to expanding the shelf life of food products.

The present review paper focuses on recent developments in edible films and coatings made of base compounds from biological sources, namely plants, animals, algae, and microorganisms. These sources include by-products, residues, and wastes from agro-food industries and sea products that contribute to sustainability concerns. Chitosan, derived from animal biological sources, such as crustacean exoskeletons, has been the most studied base compound over the past three years. Polysaccharides typically constitute no more than 3–5% of the film/coating base solution, with some exceptions, like Arabic gum. Proteins and lipids may be present in higher concentrations, such as zein and beeswax. This review also discusses the enrichment of these bio-based films and coatings with various functional and/or bioactive compounds to confer or enhance their functionalities, such as antimicrobial, antioxidant, and anti-enzymatic properties, as well as physical properties. Whenever possible, a comparative analysis among different formulations was performed. The results of the applications of these edible films and coatings to fruit and vegetable products are also described, including shelf life extension, inhibition of microbial growth, and prevention of oxidation. This review also explores novel types of packaging, such as active and intelligent packaging. The potential health benefits of edible films and coatings, as well as the biodegradability of films, are also discussed. Finally, this review addresses recent innovations in the edible films and coatings industry, including the use of nanotechnologies, aerogels, and probiotics, and provides future perspectives and the challenges that the sector is facing.

The joining of aluminium alloy AA6082-T6 to polyamide 6 (PA6) by friction stir spot welding (FSSW) was investigated in the current work. Although previous studies can be found on the joining of polymers and metals by FSSW, welding using aluminium plates as thin as the ones used in this work (1 mm) was not found. The influence of the plunge depth (0.1 to 0.5 mm) and the dwell time (15 and 30 s) parameters on the welding results was studied. In general, the increase of these parameters led to the improvement of the maximum load of the joints under tensile-shear testing. Additionally, the feasibility of multiple spot welding was tested and proven. Finally, although most of the welds were performed with a pinless tool, a tool with a conical pin and a concave shoulder was used for comparison. The use of this more conventional tool resulted in joints easily broken by handling. Still, the potential of the conical pin tool was demonstrated. The different conditions were evaluated based on morphology and tensile-shear testing. The weld with the best mechanical behaviour was produced with multiple spot welding, which failed for a maximum load of about 2350 N.

Currently, fertilization decisions in apple orchards are based on soil and leaf analyses while the leaf material is sampled after the growing season, usually in June–July (90–110 days after full bloom). This approach is inefficient, as the information becomes available later than the growing season and is therefore only useful in supporting fertilization decisions for the next year, not the current one. To establish a method that provides useful information for fertilization decisions earlier in the growth cycle, our research focused on the assessment of the nutrient content of petiole sap using different methods, the standard method and the rapid method using a reflectometer. For this study, in 2021, four ‘Gala’ orchards were fertilized with different N–P–K levels. Macro and micronutrients were determined in leaves and petiole sap at 45 and 90–110 days after full bloom (DAFB) using standard laboratory methods and the reflectometer. When leaf analysis at 45 and 90–110 DAFB was compared with petiole sap analysis at the same time point, no significant correlations were found between the nutrient contents in leaf material and petiole sap, with the exception of calcium. However, positive results were obtained regarding the correlation between reflectometer determination and standard laboratory analyses. The regression analysis revealed high determination coefficients for N-NO3− (R2 = 0.703), K+ (R2 = 0.705), Ca2+ (R2 = 0.715), and Mg2+ (R2 = 0.780) between standard laboratory methods and the reflectometer. These results suggest that the reflectometer enables a real-time diagnostic tool for monitoring nutrient status throughout the growth cycle, particularly key nutrients related to fruit quality. The N–P–K fertilization strategies had no influence on the nutrient content of leaves or petiole sap. The nutrient content of both sample types varied depending on the orchard.

Abstract Background Soil microbiomes are important to maintain soil processes in forests and confer protection to plants against abiotic and biotic stresses. These microbiomes can be affected by environmental changes. In this work, soil microbial communities from different cork oak Portuguese forests under different edaphoclimatic conditions were described by using a metabarcoding strategy targeting ITS2 and 16S barcodes. Results A total of 11,974 fungal and 12,010 bacterial amplicon sequence variants (ASVs) were obtained, revealing rich and diverse microbial communities associated with different cork oak forests. Bioclimate was described as the major factor influencing variability in these communities (or bioclimates/cork oak forest for fungal community), followed by boron and granulometry. Also, pH explained variation of fungal communities, while C:N ratio contributed to bacterial variation. Fungal and bacterial biomarker genera for specific bioclimates were described. Their co-occurrence network revealed the existence of a complex and delicate balance among microbial communities. Conclusions The findings revealed that bacterial communities are more likely to be affected by different edaphoclimatic conditions than fungal communities, also predicting a higher impact of climate change on bacterial communities. The integration of cork oak fungal and bacterial microbiota under different bioclimates could be further explored to provide information about useful interactions for increasing cork oak forest sustainability in a world subject to climate changes.