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- 2024 Annual meeting of the international network on ectopic calcification (INTEC)—abstract proceedingsPublication . Cancela, M. Leonor; Alouane, Ahmed; Bertelli, Pietro M.; Camacho, Antonio; Derudder, Robbe; Forlino, Antonella; Harris, Matthew P.; Jacinto, Marta; Lengyel, Imre; Link, Wolfgang; Murshed, Monzur; Pasch, Andreas; Kaliya-Perumal, Arun-Kumar; Quaglino, Daniela; Qin, Zihan; Sabbagh, Yves; Seminari, Elena; Villar, Marcos M.; Winkler, Christoph; Vanakker, Olivier M.The 3rd Annual Meeting of the International Network on Ectopic Calcification (INTEC) was held in Faro, Portugal on 12–13 September 2024. This hybrid meeting brought together researchers and clinicians focused on the molecular, (patho)physiological, and clinical aspects of ectopic calcification in hereditary and acquired conditions, as well as in aging. The findings presented in this year’s meeting emphasised the complexity of the field, offering new insights into both mechanistic pathways and translational hurdles. The abstracts of this year’s meeting are collected in this conference paper, with permission from the corresponding authors.
- Advanced nanotherapeutic strategies transforming diabetic wound healingPublication . Ramos, Filipa; Kumar, Girish; Virmani, Tarun; Sharma, Abhishek; Duarte, Sofia O. D.; Fonte, PedroDue to their high recurrence rates and slow healing, diabetic wounds are becoming a greater public health concern [Citation1]. Each year, 1.6 million cases of diabetic wounds occur in the United States alone, affecting approximately 18.6 million people worldwide [Citation2]. Because of poor cellular regeneration, increased inflammation, and reduced angiogenesis, traditional treatments like debridement, antibiotics, and dressings usually do not work [Citation3]. To overcome the limitations of traditional treatments, there is now a significant demand for advanced therapeutic modalities that promise accurate, efficient, and rapid healing processes [Citation4]. These include microneedles (MNs), exosomes, tetrahedral framework nucleic acids (tFNAs), three-dimensional scaffolds, gene therapy, oxygen-releasing biomaterials, phototherapies, and nanozymes.
- Aglomerular renal function in teleosts: A comparative molecular and physiological approachPublication . Pinto Teixeira, José Pedro; Pinto, Bernardo; Wilson, Jonathan M.; Guerreiro, Pedro Miguel; Castro, FilipeThe aglomerular kidney, characterised by the absence of functional glomeruli and reliance on tubular secretion alone, has evolved independently across multiple teleost lineages occupying diverse environments, including notothenioids of the Southern Ocean, gadids of cold North Atlantic and Arctic waters, and syngnathids distributed across temperate and tropical seas. Despite sharing this independently derived renal architecture, these groups face markedly distinct osmotic, thermal and chemical challenges in their natural habitats. How aglomerular kidneys cope with environmental stressors, including fluctuations in temperature and salinity, and how they handle the excretion of xenobiotics and other exogenous compounds through exclusively tubular mechanisms, remains poorly understood. The physiological and molecular responses underlying renal function in these lineages have received comparatively little attention relative to their glomerular counterparts. Objective: This study investigates how aglomerular kidneys across phylogenetically distinct teleost lineages respond, at molecular and physiological levels, to contrasting environmental conditions, namely temperature and salinity challenges predicted to alter the functional demands on renal osmoregulation. Methodology: Adult and juvenile specimens from target lineages were subjected to controlled exposure experiments combining different temperature and salinity regimes. Blood and urine samples were collected to assess osmolality and ionic composition. Renal tissues were processed for gene expression analysis of key transport and structural genes, histology, immunohistochemistry and enzymatic activity assays, providing an integrated picture of renal function under each condition. Results: Molecular analyses are currently underway. Preliminary work has established protocols for RNA extraction and quantitative PCR across target species, enabling comparative gene expression profiling to proceed across the full dataset. Conclusions: A comparative physiological and molecular approach across aglomerular teleost lineages will clarify whether shared renal architecture translates into shared functional responses to environmental challenge and identify lineage-specific mechanisms of renal adaptation, with broader relevance for predicting how these fishes may respond to ongoing environmental change.
- Antibacterial activity of PDDA-stabilized GO-AgNP nanocompositesPublication . Sadoq, Badr-Edine; Elamine, Youssef; Leal, Inês; Bouajaj, Adel; Britel, Mohammed Reda; Maurady, Amal; Power, Deborah MaryGraphene oxide (GO)-silver nanoparticle (AgNP) nanocomposites are widely studied for their antimicrobial synergy. In this study, the antibacterial activity of GO– AgNP nanocomposites prepared with poly(diallyldimethylammonium chloride) (PDDA) as a stabilizing linker was investigated. The composites were characterized by UV–Vis spectroscopy, FTIR, zeta potential analysis, XRD, TGA, and MPAES. GO–PDDA–AgNPs exhibited a strong surface plasmon resonance band at ~ 420 nm, new C–H stretching bands at 2865–3012 cm⁻¹ from PDDA, and a high positive zeta potential (+ 57.5 mV) compared to bare GO (–40 mV), confirming successful functionalization and improved colloidal stability. Antibacterial activity was evaluated against E. coli and S. aureus using disk diffusion assays, growth curves, and MIC determination. GO and AgNPs alone showed no inhibition at concentrations up to 20 µg/mL, whereas PDDA alone produced inhibition zones of 6–11.5 mm against S. aureus (2.5–100 µg/mL) and induced selective bacterial aggregation. The minimum inhibitory concentration (MIC) of PDDA was between 6.25 and 12.5 µg/mL for S. aureus and between 25 and 50 µg/mL for E. coli, indicating greater potency toward Gram-positive bacteria. The GO–PDDA–AgNP composite inhibited S. aureus with zones of 8.5–10 mm at 10–20 µg/mL, while no significant inhibition was observed for E. coli at the tested concentrations. Molecular docking simulations examining interactions between PDDA and quorum-sensing regulatory proteins AgrA in S. aureus and LsrR in E. coli predicted higher binding affinity to AgrA (–5.11 kcal/mol) than LsrR (–3.76 kcal/mol). While in vitro assays using a Chromobacterium violaceum CV026 biosensor showed no inhibition of AHL-mediated signalling, it should be noted that this model differs mechanistically from the Gram-positive agr system, leaving the predicted AgrA interaction as a potential target for future investigation. Ultimately, this study demonstrated that PDDA is the primary antibacterial component of the GO–PDDA–AgNP composite, exhibiting potent activity against Gram-positive bacteria through a mechanism involving selective bacterial aggregation.
- Antibody–drug conjugates: pharmacotherapeutic properties and future perspectivesPublication . Augusto, André; Cristiano, Maria de Lurdes; Conceição, JaimeBackground: The clinical landscape for antibody–drug conjugates (ADCs) is currently experiencing an unprecedented expansion, with more than 20 agents approved to date and hundreds presently under clinical evaluation, underscoring their growing impact in precision oncology. By combining the cytotoxic potency of chemotherapy with the selectivity of monoclonal antibodies, ADCs have redefined targeted cancer therapy. Nevertheless, challenges related to toxicity, resistance, and suboptimal drug delivery continue to limit their full clinical potential. Objectives: This review provides a comprehensive description of currently approved ADCs, with a particular focus on their pharmacotherapeutic properties, mechanisms of action, therapeutic indications, and safety profiles. By integrating currently available clinical data and pharmacological properties, it is possible to identify key translational gaps between ADC design and their real-world performance. This article also evaluates how the structural components contribute to both efficacy and toxicity of ADCs, offering a framework for rational molecular optimizations. Conclusions: Beyond the current oncology-centric paradigm, this review highlights the imminent pivot toward non-oncology applications, including targeted therapies for autoimmune, infectious, and neurodegenerative diseases. Importantly, this article highlights emerging innovations shaping the next generation of ADCs, including bispecific antibodies, novel cytotoxic payloads with improved therapeutic indices, and advanced linker technologies enabling more precise payload release. Despite current limitations, ongoing advances in ADC development, along with a rapidly expanding clinical pipeline, position these drugs in a dynamic therapeutic class with the potential to transform multiple complex diseases and improve the quality of life of patients who have them.
- Are there differential roles in the parental behaviour of the chameleon cichlid australoheros facetus?Publication . Oliveira, Gonçalo; Saraiva, João; Jesus, Nuno F.; Guerreiro, Pedro MiguelCichlids are social fishes well known for their complex behaviour. The chameleon cichlid, Autraloheros facetus, is native to South American river drainages and is currently established in several Mediterranean-type drainages in southern Portugal as an invasive species. Their high local recruitment, territorially, and parental care activities are possible advantages in competing with native fish and achieving high reproductive success. The main objective of this work was to characterise the behaviours of the males and females of the species A. facetus upon pair formation and their roles during parental behaviour at different stages of offspring development for the purpose of gathering important basic knowledge on fish biology to apply in the control of the species’ populations. To attain this initial goal, we used observation techniques and video recording protocols to characterise the specific activities performed by each individual during reproductive and parental stages (pair formation, eggs, attached larvae, and free-swimming larvae), identifying the main tasks, and assessing the time spent on each task by each member of the reproductive pair. The breeding pairs were obtained as a result of the social hierarchy formed in each social group: groups of six individuals of similar size were placed in individual tanks fitted with a bottom biological filter, in which the temperature was increased to 24 ◦C at an expanded photoperiod. Social behaviours were recorded and characterised, expanded on a previously established ethogram. The results obtained so far allow us to establish a set of aggressive behaviours towards other fish (striking, chasing, biting), nest preparing behaviours (digging, cleaning), caring (caring, fetching), and guarding behaviours towards the offspring (hovering, patrolling). Concerning the rate of occurrence of social behaviours in randomised 5 min periods (frequency of specific behaviours per each 5 min period) show that in the pair formation stage, males present a more aggressive nature with frequent occurrence of striking (rstriking = 1) and biting (rbiting = 0.31). On the other hand, at the egg stage, the rate of occurrence of parental behaviours shows the dominance of females to prepare the nest (rdigging = 0.91) and caring for the eggs (rcaring = 3.64), while males are more vigilant (rparental hovering = 0.95). At the attached larvae stage, this tendency continues with higher occurrence rates: males focus on vigilance (rparental hovering = 1.6) and patrolling (rpatrolling = 2), and females care for the recently hatched larvae (rfetching = 4). Finally, at the free-swimming larvae stage, there is a turn and a small decrease in the occurrence of these activities: males oversee nest maintenance (rdigging = 1.09), and females patrol the tank (rpatrolling = 0.71) and care for the larvae (rfetching = 2.02). These preliminary data suggest differential roles for male and female A. facetus, that evolve during parental behaviour, for which further experimental paradigms will be designed to explore underlying proximate causes.
- Assessing the bioactive potential of Lysimachia atropurpurea extracts using HPLC-MS/MS, in vitro and in silico analysisPublication . Ak, Gunes; Nilofar, Nilofar; Saka, Enver; Uba, Abdullahi Ibrahim; Rodrigues, Maria João; Fernandes, Eliana; Custódio, Luísa; Yildiztugay, Evren; Yapıcı, Ismail; Gulcin, Ilhami; Mahmoud, Orchid A.; Eldahshan, Omayma A.; Singab, Abdel Nasser B.; Wu, Yimao; Li, Meng-Yao; Zengin, GokhanThe genus Lysimachia is of great interest to the scientific community, especially in terms of its potential anticancer effects. In this study, the aerial parts and roots of Lysimachia atropurpurea L. were collected and extracted by maceration using solvents of ethyl acetate (EA), ethanol (EtOH), ethanol/water, and water. The biological activities of the extracts, including antioxidant, enzyme inhibition, and anticancer effects, were evaluated using various assays. High-performance liquid chromatographytandem mass spectrometry (HPLC-MS/MS) analysis revealed a total of 32 compounds in the extracts of L. atropurpurea. The roots showed significantly the highest antioxidant activity compared to the aerial part. In case of cholinesterase inhibition, the aerial parts of the EtOH extract showed the highest acetylcholinesterase (AChE) inhibition activity, measuring 3.05 mg galatamine equivalent (GALAE)/g. The EtOH and EtOH/water extracts exhibited the strongest cytotoxicity, reducing the viability of human neuroblastoma (SH-SY5Y) and human hepatocarcinoma (HepG2) cancer cells to as low as 4.86–6.33 %. The results of network pharmacology and molecular docking suggest that the extract of L. atropurpurea exerts inhibitory effects on hepatocellular carcinoma through the modulation of SRC, PI3K, and HSP90, while it demonstrates potential inhibitory activity against neuroblastoma by targeting SRC, PI3K, HSP90, ESR1, AKT, and other related targets. In conclusion, the L. atropurpurea extracts showed potential antioxidant, enzyme inhibition, and selective anticancer effects, which support their potential for further research as therapeutic agents in drug development.
- Assessment of cell disruption methods in an integrated multi-product biorefinery for Nannochloropsis Oceanica: from process design to economic analysis.Publication . Cunha, Pedro; Carvalho, Bernardo; Kholany, Mariam; Pereira, Hugo; Varela, JoãoMicroalgae are bioresources with significant potential within a sustainable, circular, bio-economy. However, high production costs have limited the widespread use of algae biomass. This study aimed to develop a multi-product biorefinery for Nannochloropsis oceanica that generates multiple revenue streams from the biomass, thereby enhancing the economic viability of algal production. The effectiveness of cell wall disruption using high-pressure homogenization and enzymatic hydrolysis was evaluated. Enzymatic hydrolysis solubilized nearly half (48.2 ± 1.5%) of the dry cell weight, compared to only 27.3 ± 3.2% with high-pressure homogenization, resulting in more concentrated water-soluble fractions and significantly higher protein extraction yields. Lipid extracts obtained after enzymatic hydrolysis had higher lipid (72.0 ± 5.3% w/w) and eicosapentaenoic acid (28.1 ± 6.9% w/w) contents than those from high-pressure homogenization (38.8 ± 6.1% w/w lipids; 9.1 ± 0.6% w/w eicosapentaenoic acid), despite similar lipid extraction yields (around 30%). Increasing the ethanol volumetric ratio from 58% to 75% v/v significantly improved lipid extraction yields (57.4 ± 3.1%) in the enzymatic hydrolysis-based biorefinery, with even higher yields observed upon scaling up (70.1%). All fractions, including lipid extracts, exhibited a balanced essential amino acid profile that exceeded the WHO/FAO/UNU-recommended values. A preliminary economic analysis indicated that lipid production was more cost-effective when cells were permeabilized by enzymatic hydrolysis than by high-pressure homogenization.
- Assessment of the reproductive status of captive populations of endangered leuciscid species from the iberian peninsula: a. hispanica, I. lusitanicum and a. occidentalePublication . Hernandez, Ana; Gil, Fátima; Sousa-Santos, Carla; Cabrita, Elsa; Guerreiro, Pedro Miguel; Gallego, VictorPopulations of freshwater fish species endemic to the Iberian Peninsula have been declining since the mid-20th century, and the captive breeding of highly endangered species is considered to be a useful tool to restock declining populations. A pioneer project of supportive breeding of critically endangered fish started in 2007 at the Aquário Vasco da Gama (AVG), and this work aims to show the reproductive status of the breeders which make up the current captive broodstoks. Populations of different endangered leuciscid species (Anaecypris hispanica, Iberochondrostoma lusitanicum and Achondrostoma occidentale) were sampled at AVG during the spring of 2022. Breeders were counted and sexed, and males were stripped to check for the presence of spermatozoa. The sperm volume was assessed visually, and spermatozoa motility was assessed by a CASA system. Sperm samples were classified into four classes based on the percentage of motile cells: C-I ≤ 25%, C-II = 25–50%, C-III = 50–75%; and C-IV > 75%. The captive population of A. hispanica consisted of 63 individuals and showed a 40% of spermiating males, with an average volume of 5–10 µL. The histogram of sperm quality reported that 15% males had sperm motility of C-II, 50% of males had sperm motility of C-III and, finally, 35% of males had sperm with the high-quality class (C-IV). The population of I. lusitanicum consisted of 599 individuals and showed 93% of spermiating males, with an average volume of 15–20 µL. The histogram of sperm quality reported that most part of the males had good sperm quality belonging to C-III and C-IV class (26% and 71%, respectively), while just 1 male showed bad quality sperm (C-II). The captive population of A. occidentale consisted of 193 individuals, showing a 62% of spermiating males with an average volume of 20–25 µL. The histogram showed that 6% males had sperm motility of C-I, 26% of males had sperm motility of C-II, the most part of the males (45%) showed a sperm quality of C-III and, finally, 23% of males had sperm with the high-quality class (C-IV). Since the project began in 2007, more than 12,000 fish of these three critically endangered species have been released to restock the populations from which the respective wild breeders were caught. All captive fish were released after a maximum of three consecutive generations in captivity, and new stocks were established with wild adults from the target populations, to avoid the negative effects of inbreeding and genetic drift on the original genetic pool.
- Atlantic arc lander monitoring (ALaMo): an emerging network of low-cost lander arrays for ocean bottom observationsPublication . González-Pola, Cesar; Cusack, Caroline; Urquijo, Ignacio; Graña, Rocio; Rodriguez-Cobo, Luis; Sánchez-Leal, Ricardo; Nolan, Glenn; Santos, A. Miguel P.The need to establish a sustained Global Ocean Observing System (GOOS) has long been recognized by the international ocean science community. Established in 1991 and led by the Intergovernmental Oceanographic Commission of the United Nations Educational, Scientific, and Cultural Organization (IOC-UNESCO), GOOS develops guidelines and coordinates regional alliances across the world’s ocean basins to evolve the system. The need to sustainably expand GOOS has recently gained urgency as expressed in the United Nations Decade of Ocean Science for Sustainable Development (2021–2030) Challenge 7: Ensure a sustainable ocean observing system across all ocean basins that delivers accessible, timely, and actionable data and information to all users.