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Mycobacterium tuberculosis (Mtb), the aetiologic agent of tuberculosis (TB), stores triacylglycerol (TAG) in the form of intrabacterial lipid inclusions (ILI) to survive and chronically persist within its host. These highly energetic molecules represent a major source of carbon to support bacterial persistence and reactivation, thus playing a leading role in TB pathogenesis. However, despite its physiological and clinical relevance, ILI metabolism in Mtb remains poorly understood. Recent discoveries have suggested that several ILI‐associated proteins might be widely conserved across TAG‐producing prokaryotes, but still very little is known regarding the nature and the biological functions of these proteins. Herein, we performed an in silico analysis of three independent ILI‐associated proteomes previously reported to computationally define a potential core ILI‐associated proteome, referred to as ILIome. Our investigation revealed the presence of 70 orthologous proteins that were strictly conserved, thereby defining a minimal ILIome core. We further narrowed our analysis to proteins involved in lipid metabolism and discuss here their putative biological functions, along with their molecular interactions and dynamics at the surface of these bacterial organelles. We also highlight the experimental limitations of the original proteomic investigations and of the present bioinformatic analysis, while describing new technological approaches and presenting biological perspectives in the field. The in silico investigation presented here aims at providing useful datasets that could constitute a scientific resource of broad interest for the mycobacterial community, with the ultimate goal of enlightening ILI metabolism in prokaryotes with a special emphasis on Mtb pathogenesis.

Integrating Artificial Intelligence (AI) into Precision Medicine (PM) is redefining healthcare, enabling personalized treatments tailored to individual patients based on their genetic code, environment, and lifestyle. AI’s ability to analyze vast and complex datasets, including genomics and medical records, facilitates the identification of hidden patterns and correlations, which are critical for developing personalized treatment plans. Unsupervised Learning (UL) is particularly valuable in PM as it can analyze unstructured and unlabeled data to uncover novel disease subtypes, biomarkers, and patient stratifications. By revealing patterns that are not explicitly labeled, unsupervised algorithms enable the discovery of new insights into disease mechanisms and patient variability, advancing our understanding of individual responses to treatment. However, the integration of AI into PM presents some challenges, including concerns about data privacy and the rigorous validation of AI models in clinical practice. Despite these challenges, AI holds immense potential to revolutionize PM, offering a more personalized, efficient, and effective approach to healthcare. Collaboration among AI developers and clinicians is essential to fully realize this potential and ensure ethical and reliable implementation in medical practice. This review will explore the latest emerging UL technologies in the biomedical field with a particular focus on PM applications and their impact on human health and well-being.

Currently, one-third of global food production, accounting for 1.3 billions tons, goes wasted due to major humanitarian and environmental challenges. In such a scenario, the circular bioeconomy model stands as an innovative solution by promoting sustainable production, utilizing agri-food waste, and converting non-renewable products into valuable resources. Here, the circular bioeconomy concept was applied on a previously obtained chestnut burr extract (agri-food waste) composed of gallic acid, quinic acid, protocatechuic acid, brevifolin carboxylic acid, and ellagic acid to evaluate its antimicrobial activity against four bacterial opportunistic pathogens (Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli). Our results evidenced a modest but measurable antibacterial activity against Enterococcus faecalis, with a minimum inhibitory concentration (MIC) of 64 μg/mL. In silico studies allowed for identifying the potential molecular target, supporting the underlying antibacterial activity of the active principle and providing useful molecular findings regarding their interaction. In this study, we show a robust and comprehensive in vitro and in silico pipeline aimed at the identification of novel antibacterial scaffolds taking advantage of agri-food waste.

Due to the rising annual incidence of lung cancer (LC), new treatment strategies are needed. While various options exist, many, if not all, remain suboptimal. Several studies have shown cryoablation to be a promising approach. Yet, a lack of basic information pertaining to LC response to freezing and requirement for percutaneous access has limited clinical use. In this study, we investigated the A549 lung carcinoma cell line response to freezing. The data show that a single 5 min freeze to −15 °C did not affect cell viability, whereas −20 °C and −25 °C result in a significant reduction in viability 1 day post freeze to

Intracellular pathogens such as Mycobacterium tuberculosis (Mtb) have evolved diverse strategies to counteract macrophage defence mechanisms including phagolysosomal biogenesis. Within macrophages, Mtb initially resides inside membrane‐bound phagosomes that interact with lysosomes and become acidified. The ability of Mtb to control and subvert the fusion between phagosomes and lysosomes plays a key role in the pathogenesis of tuberculosis. Therefore, understanding how pathogens interact with the endolysosomal network and cope with intracellular acidification is important to better understand the disease. Here, we describe in detail the use of fluorescence microscopy‐based approaches to investigate Mtb responses to acidic environments in cellulo. We report high‐content imaging modalities to probe Mtb sensing of external pH or visualise in real‐time Mtb intrabacterial pH within infected human macrophages. We discuss various methodologies with step‐by‐step analyses that enable robust image‐based quantifications. Finally, we highlight the advantages and limitations of these different approaches and discuss potential alternatives that can be applied to further investigate Mtb–host cell interactions. These methods can be adapted to study host–pathogen interactions in different biological systems and experimental settings. Altogether, these approaches represent a valuable tool to further broaden our understanding of the cellular and molecular mechanisms underlying intracellular pathogen survival.

The enzyme 4-hydroxyphenylpyruvate dioxygenase (4-HPPD) is involved in the catabolism of the amino acid tyrosine in organisms such as bacteria, plants, and animals. It catalyzes the conversion of 4-hydroxyphenylpyruvate to a homogenisate in the presence of molecular oxygen and Fe(II) as a cofactor. This enzyme represents a key step in the biosynthesis of important compounds, and its activity deficiency leads to severe, rare autosomal recessive disorders, like tyrosinemia type III and hawkinsinuria, for which no cure is currently available. The 4-HPPD C-terminal tail plays a crucial role in the enzyme catalysis/gating mechanism, ensuring the integrity of the active site for catalysis through fine regulation of the C-terminal tail conformation. However, despite growing interest in the 4-HPPD catalytic mechanism and structure, the gating mechanism remains unclear. Furthermore, the absence of the whole 3D structure makes the bioinformatic approach the only possible study to define the enzyme structure/molecular mechanism. Here, wild-type 4-HPPD and its mutants were deeply dissected by applying a comprehensive bioinformatics/evolution study, and we showed for the first time the entire molecular mechanism and regulation of the enzyme gating process, proposing the full-length 3D structure of human 4-HPPD and two novel key residues involved in the 4-HPPD C-terminal tail conformational change.

Utilizing plant-based resources, particularly their by-products, aligns with sustainability principles and circular bioeconomy, contributing to environmental preservation. The therapeutic potential of plant extracts is garnering increasing interest, and this study aimed to demonstrate promising outcomes from an extract obtained from an underutilized plant waste. Chaetomorpha linum, an invasive macroalga found in the Orbetello Lagoon, thrives in eutrophic conditions, forming persistent mats covering approximately 400 hectares since 2005. The biomass of C. linum undergoes mechanical harvesting and is treated as waste, requiring significant human efforts and economic resources—A critical concern for municipalities. Despite posing challenges to local ecosystems, the study identified C. linum as a natural source of bioactive metabolites. Phytochemical characterization revealed lipids, amino acids, and other compounds with potential anti-inflammatory activity in C. linum extract. In vitro assays with LPS-stimulated RAW 264.7 and TNF-α/IFN-γ-stimulated HaCaT cells showed the extract inhibited reactive oxygen species (ROS), nitric oxide (NO), and prostaglandin E2 (PGE2) productions, and reduced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expressions via NF-κB nuclear translocation, in RAW 264.7 cells. It also reduced chemokines (TARC/CCL17, RANTES/CCL5, MCP-1/CCL2, and IL-8) and the cytokine IL-1β production in HaCaT cells, suggesting potential as a therapeutic candidate for chronic diseases like atopic dermatitis. Finally, in silico studies indicated palmitic acid as a significant contributor to the observed effect. This research not only uncovered the untapped potential of C. linum but also laid the foundation for its integration into the circular bioeconomy, promoting sustainable practices, and innovative applications across various industries.

Despite cardiac magnetic resonance (CMR) with late gadolinium enhancement (LGE) being the current gold standard for non-invasive myocardial characterization and fibrosis quantification, its accessibility is limited, particularly in acute settings and in certain patient populations with contraindications to magnetic resonance imaging. Late iodine enhancement (LIE) in computed tomography (CT) imaging has emerged as a potential alternative, capitalizing on the similarities in the contrast kinetics between gadolinium and iodinated contrast agents. Studies have investigated LIE-CT’s effectiveness in myocardial infarction (MI) detection, revealing promising outcomes alongside some disparities compared to LGE-CMR. LIE-CT also proves beneficial in diagnosing non-ischemic heart diseases such as myocarditis, hypertrophic cardiomyopathy, and sarcoidosis. While LIE-CT demonstrates good accuracy in detecting certain myocardial pathologies, including acute MI and chronic fibrotic changes, it has limitations, such as the inability to detect diffuse myocardial enhancement. Nonetheless, thanks to the availability of optimized protocols with minimal radiation doses and contrast medium administration, integrating LIE-CT into cardiac CT protocols could enhance its clinical utility, particularly in acute settings, providing valuable prognostic and management insights across a spectrum of cardiac ischemic and non-ischemic conditions.

This retrospective study aims to further investigate the effects of rapid maxillary expansion (RME) treatment on respiratory function in growing patients with contracted upper airways, by assessing nasal resistance using active anterior rhinomanometry (AAR). Methods: At T0 (pre-RME), 122 orthodontic patients aged 5 to 12 years, exhibiting oral breathing and/or snoring along with maxillary contraction, underwent AAR with a mask using the ATMOS Rhino 31 rhinomanometer. Additionally, the SRDB or PSQ-SRDB Questionnaire was utilized to validate the orthodontic history of obstructive sleep apnea syndrome (OSAS). If AAR results at T0 were clinically significant, they were repeated at T1, corresponding to a period of three months after screw fixation, to evaluate changes in nasal resistance values between pre- and post-treatment. Results: The study group comprised 42 oral respiratory patients with a mean age of 7.71 years, suspected of pediatric OSAS. Although the questionnaires did not yield statistically significant results, the therapy induced an overall reduction in airflow resistance values from both nostrils (−24.63% in the right nostril; −26.65% in the left nostril). Conclusions: This study demonstrates the beneficial effects of orthodontic treatment with RME on nasal function in terms of airflow resistance in cases of maxillary contraction.

A. baumannii can rapidly acquire new resistance mechanisms and persist on abiotic surface, enabling the colonization of asymptomatic human host. In Acinetobacter the type VI secretion system (T6SS) is involved in twitching, surface motility and is used for interbacterial competition allowing the bacteria to uptake DNA. A. baumannii possesses a T6SS that has been well studied for its regulation and specific activity, but little is known concerning its assembly and architecture. The T6SS nanomachine is built from three architectural sub-complexes. Unlike the baseplate (BP) and the tail-tube complex (TTC), which are inherited from bacteriophages, the membrane complex (MC) originates from bacteria. The MC is the most external part of the T6SS and, as such, is subjected to evolution and adaptation. One unanswered question on the MC is how such a gigantesque molecular edifice is inserted and crosses the bacterial cell envelope. The A. baumannii MC lacks an essential component, the TssJ lipoprotein, which anchors the MC to the outer membrane. In this work, we studied how A. baumannii compensates the absence of a TssJ. We have characterized for the first time the A. baumannii's specific T6SS MC, its unique characteristic, its membrane localization, and assembly dynamics. We also defined its composition, demonstrating that its biogenesis employs three Acinetobacter-specific envelope-associated proteins that define an intricate network leading to the assembly of a five-proteins membrane super-complex. Our data suggest that A. baumannii has divided the function of TssJ by (1) co-opting a new protein TsmK that stabilizes the MC and by (2) evolving a new domain in TssM for homo-oligomerization, a prerequisite to build the T6SS channel. We believe that the atypical species-specific features we report in this study will have profound implication in our understanding of the assembly and evolutionary diversity of different T6SSs, that warrants future investigation.

Encephalic vascular accident, or stroke, is the most common pathology of the central nervous system in humans, the second leading cause of death and physical and cognitive disabilities, in developing countries. It presents as an ischemic (more common) or hemorrhagic form. Ozone therapy has been shown to be effective in neuromodulation, neuroprotection, and nerve regeneration. The present study aimed to evaluate the effect of targeted mild ozone after inducing cerebral ischemia in vitro. Neuroblastoma lineage cells (SH-SY5Y) and canine amniotic membrane stem cells were subjected to 24 hours of hypoxia in an incubator culture chamber. The cells were evaluated by MTT assay, colorimetric assay spectrophotometry, fluorescence microscopy, and flow cytometry. Treatment with low concentrations of ozone (2–10 µg/mL), indicated a possible neuroregenerative effect at low concentrations, correlated with lower levels of apoptosis and oxidative stress compared to cells not subjected to hypoxia. High concentrations of ozone (18–30 µg/mL) promoted an increase in rate of apoptosis and cell death. We developed a novel protocol that mimics ozone therapy for ischemic stroke, using ozonized culture medium after hypoxia induction. Although more studies are needed, we conclude that ozone has a dose-dependent hormetic effect and can reverse the effect of ischemia in vitro at low concentrations.

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease that may be treated utilizing thermal therapies. Cryoablation is an effective, minimally invasive therapy that has been utilized for the treatment of various cancers, offering patients a quicker recovery and reduced side effects. Cryoablation has been utilized on a limited basis for the treatment of PDAC. With the recent reports on the success of cryoablation, there is a growing interest in the use of cryoablation as a standalone, minimally invasive procedure to treat PDAC. While offering a promising path, the application of cryoablation to PDAC is limited by current technologies. As such, there is a need for the development of new devices to support advanced treatment strategies for PDAC. To this end, this study investigated the performance of a new endoscopic ultrasound-compatible cryoablation catheter technology, FrostBite. We hypothesized that FrostBite would enable the rapid, effective, minimally invasive delivery of ultra-cold temperatures to target tissues, resulting in effective ablation via an endoscopic approach. Thermal properties and ablative efficacy were evaluated using a heat-loaded gel model, tissue-engineered models (TEMs), and an initial in vivo porcine study. Freeze protocols evaluated included single and repeat 3 and 5 min applications. Isotherm assessment revealed the generation of a 2.2 cm diameter frozen mass with the −20 °C isotherm reaching a diameter of 1.5 cm following a single 5 min freeze. TEM studies revealed the achievement of temperatures ≤ −20 °C at a diameter of 1.9 cm after a 5 min freeze. Fluorescent imaging conducted 24 h post-thaw demonstrated a uniformly shaped ellipsoidal ablative zone with a midline diameter of 2.5 cm, resulting in a total ablative volume of 6.9 cm3 after a single 5 min freeze. In vivo findings consistently demonstrated the generation of ablative areas measuring 2.03 cm × 3.2 cm. These studies demonstrate the potential of the FrostBite cryocatheter as an endoscopic ultrasound-based treatment option. The data suggest that FrostBite may provide for the rapid, effective, controllable freezing of cancerous pancreatic and liver tissues. This ablative power also offers the potential of improved safety margins via the minimally invasive nature of an endoscopic ultrasound-based approach or natural orifice transluminal endoscopic surgery (NOTES)-based approach. The results of this pre-clinical feasibility study show promise, affirming the need for further investigation into the potential of the FrostBite cryocatheter as an advanced, minimally invasive cryoablative technology.

Tritium permeation constitutes a key issue for the future EU-DEMO, especially in the Breeding Blanket (BB) where fusion energy must be delivered to the Primary Heat Transport System (PHTS) and where tritium must be bred. Currently, the mitigation strategy of the tritium permeation from BB into primary coolant is based on the adoption of anti-permeation barriers and on the operation of the Coolant Purification System (CPS). This system must ensure a tritium removal rate from the primary coolant equal to the BB permeation rate at a target tritium-specific activity inside the PHTS. In the case of the Water-Cooled Lithium Lead (WCLL) BB, water distillation was selected as the most promising technology for the primary coolant detritiation due to its intrinsic simplicity and safety. Nevertheless, power consumption was recognized as a relevant concern. For this reason, the present work aims at investigating possibilities to reduce power consumption of the water CPS implementing Heat Pump-Assisted Distillation (HPAD) concepts. To do this, a review of the HPADs developed in the chemical industry was carried out, and the best options for the water CPS were identified based on qualitative considerations. Then, a quantitatively assessment of the best solution in terms of power consumption and tritium inventory was performed with the commercial numerical tool Aspen Plus. Finally, the Mechanical Vapor Recompression (MVR) concept was recognized as the most promising solution, ensuring a power saving of around 80% while keeping a limited tritium inventory.

While technological advancements and their deep integration in connected and automated vehicles is a central aspect in the evolving trend of automotive industry, they also depict a growing size attack surface for malicious actors: the latter ones typically aim at exploiting known and unknown security vulnerabilities, with potentially disastrous consequences on the safety of vehicles, people, and infrastructures. In recent years, remarkable efforts have been spent to mitigate security vulnerabilities in intelligent and connected vehicles, in particular in the inside of vehicles, the so-called intra-vehicle networks. Despite those efforts, securing intra-vehicle networks remains a non-trivial task due to their heterogeneous and increasingly complex context. Starting from the above remarks and motivated by the industrial research and innovation project EMERGE, in this paper we report on a novel cryptographic hardware-software solution that we have designed and developed for securing the intra-vehicle network of intelligent connected vehicles: the Crypto-Engine. The Crypto-Engine relies on a lightweight hybrid-key cryptographic scheme to provide confidentiality and authentication without compromising the normal communication performance. We tested the Crypto-Engine and demonstrated that, once configured according to application-defined performance requirements, it can authenticate parties and secure the communications with a negligible overhead.

Alkaptonuria (AKU), a rare genetic disorder, is characterized by the accumulation of homogentisic acid (HGA) in the body. Affected individuals lack functional levels of an enzyme required to breakdown HGA. Mutations in the homogentisate 1,2-dioxygenase (HGD) gene cause AKU and they are responsible for deficient levels of functional HGD, which, in turn, leads to excess levels of HGA. Although HGA is rapidly cleared from the body by the kidneys, in the long term it starts accumulating in various tissues, especially cartilage. Over time (rarely before adulthood), it eventually changes the color of affected tissue to slate blue or black. Here we report a comprehensive mutation analysis of 111 pathogenic and 190 non-pathogenic HGD missense mutations using protein structural information. Using our comprehensive suite of graph-based signature methods, mCSM complemented with sequence-based tools, we studied the functional and molecular consequences of each mutation on protein stability, interaction and evolutionary conservation. The scores generated from the structure and sequence-based tools were used to train a supervised machine learning algorithm with 89% accuracy. The empirical classifier was used to generate the variant phenotype for novel HGD missense mutations. All this information is deployed as a user friendly freely available web server called HGDiscovery (https://biosig.lab.uq.edu.au/hgdiscovery/).

Drugs that target human thymidylate synthase (hTS), a dimeric enzyme, are widely used in anticancer therapy. However, treatment with classical substrate-site-directed TS inhibitors induces over-expression of this protein and development of drug resistance. We thus pursued an alternative strategy that led us to the discovery of TS-dimer destabilizers. These compounds bind at the monomer-monomer interface and shift the dimerization equilibrium of both the recombinant and the intracellular protein toward the inactive monomers. A structural, spectroscopic, and kinetic investigation has provided evidence and quantitative information on the effects of the interaction of these small molecules with hTS. Focusing on the best among them, E7, we have shown that it inhibits hTS in cancer cells and accelerates its proteasomal degradation, thus causing a decrease in the enzyme intracellular level. E7 also showed a superior anticancer profile to fluorouracil in a mouse model of human pancreatic and ovarian cancer. Thus, over sixty years after the discovery of the first TS prodrug inhibitor, fluorouracil, E7 breaks the link between TS inhibition and enhanced expression in response, providing a strategy to fight drug-resistant cancers.

Abstract Objective Children with functional abdominal pain disorders (FAPD) and clinical elevations in three risk areas (anxiety, functional disability, and pain) have been found to be at increased risk for persistent disability. We evaluated if the presence of these three risk factors corresponded with greater gastrointestinal inflammation (measured via fecal calprotectin; FC) compared to those with no risk factors. FC concentration differences between children with three risk factors and those with one and two risk factors were explored. Results Fifty-six children with FAPD (M age = 12.23) completed measures of anxiety (Screen for Child Anxiety Related Disorders), disability (Functional Disability Inventory), and pain intensity (Numeric Rating Scale). Participants were stratified into risk groups (range: 0–3). Fisher’s exact tests were conducted to determine if children with three versus fewer risk factors were more likely to have elevated FC (≥ 50 µg/g) versus normal levels. Children with three risk factors (M FC = 86.04) were more likely to have elevated FC compared to children with zero (M FC = 25.78), one (M FC = 38.59), and two risk factors (M FC = 45.06; p’s

Conventional therapy options for chronic pain are still insufficient and patients most frequently request alternative medical treatments, such as medical cannabis. Although clinical evidence supports the use of cannabis for pain, very little is known about the efficacy, dosage, administration methods, or side effects of widely used and accessible cannabis products. A possible solution could be given by pharmacogenetics, with the identification of several polymorphic genes that may play a role in the pharmacodynamics and pharmacokinetics of cannabis. Based on these findings, data from patients treated with cannabis and genotyped for several candidate polymorphic genes (single-nucleotide polymorphism: SNP) were collected, integrated, and analyzed through a machine learning (ML) model to demonstrate that the reduction in pain intensity is closely related to gene polymorphisms. Starting from the patient’s data collected, the method supports the therapeutic process, avoiding ineffective results or the occurrence of side effects. Our findings suggest that ML prediction has the potential to positively influence clinical pharmacogenomics and facilitate the translation of a patient’s genomic profile into useful therapeutic knowledge.

The recent advancements in Artificial Intelligence have paved the way for remarkable achievements in tasks that have traditionally posed challenges even for humans [...]

Emerging machine learning (ML) technologies have the potential to significantly improve the research and treatment of rare diseases, which constitute a vast set of diseases that affect a small proportion of the total population. Artificial Intelligence (AI) algorithms can help to quickly identify patterns and associations that would be difficult or impossible for human analysts to detect. Predictive modeling techniques, such as deep learning, have been used to forecast the progression of rare diseases, enabling the development of more targeted treatments. Moreover, AI has also shown promise in the field of drug development for rare diseases with the identification of subpopulations of patients who may be most likely to respond to a particular drug. This review aims to highlight the achievements of AI algorithms in the study of rare diseases in the past decade and advise researchers on which methods have proven to be most effective. The review will focus on specific rare diseases, as defined by a prevalence rate that does not exceed 1–9/100,000 on Orphanet, and will examine which AI methods have been most successful in their study. We believe this review can guide clinicians and researchers in the successful application of ML in rare diseases.

The future of education lies in the ability to develop technologies which integrate seamless artificial intelligence (AI) components into the educational process, in order to deliver a personalized service which is dynamically tailored to the learner’s characteristics, abilities, and needs [...]

SARS–CoV-2 (COVID-19) infection can cause a severe respiratory distress syndrome. The risk of severe manifestations and mortality characteristically increase in the elderly and in the presence of non-COVID-19 comorbidity. We and others previously demonstrated that the low molecular weight (LMW) and protein thiol/disulfide ratio declines in human plasma with age and such decline is even more rapid in the case of inflammatory and premature aging diseases, which are also associated with the most severe complications of COVID-19 infection. The same decline with age of the LMW thiol/disulfide ratio observed in plasma appears to occur in the extracellular fluids of the respiratory tract and in association with many pulmonary diseases that characteristically reduce the concentrations and adaptive stress response of the lung glutathione. Early evidence in literature suggests that the thiol to disulfide balance of critical Cys residues of the COVID-19 spike protein and the ACE-2 receptor may influence the risk of infection and the severity of the disease, with a more oxidizing environment producing the worst prognosis.With this hypothesis paper we propose that the age-dependent decline of LMW thiol/disulfide ratio of the extracellular fluids, could play a role in promoting the physical (protein-protein) interaction of CoV-2 and the host cell in the airways. Therefore, this redox-dependent interaction is expected to affect the risk of severe infection in an age-dependent manner. The hypothesis can be verified in experimental models of in vitro CoV-2 infection and at the clinical level in that LMW thiols and protein thiolation can now be investigated with standardized, reliable and versatile laboratory protocols. Presenting the verification strategy of our hypothesis, we also discuss available nutritional and ancillary pharmacological strategies to intervene on the thiol/disulfide ratio of extracellular fluids of subjects at risk of infection and COVID-19 patients.

Anaemia is an important cause of morbidity and mortality globally. Among infectious agents responsible for anaemia, helminthic infections are often neglected, particularly in non-endemic countries. However, they should not be neglected in this setting, as international travel and migration are on the rise. In this narrative review, we aimed to describe soil-transmitted helminths as a cause of or contributing factor to anaemia, focusing on hookworms (Necator americanus and Ancylostoma duodenale), the whipworm (Trichuris trichiura), the roundworm (Ascaris lumbricoides), and the threadworm (Strongyloides stercoralis). A general review on the epidemiology, lifecycle, and clinical spectrum of anaemia is proposed, with a special focus on helminthic infections’ association with anaemia as well as the diagnostic approach, which are both particularly important in non-endemic settings.

Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), an infectious disease that spreads rapidly in humans. In March 2020, the World Health Organization (WHO) declared a COVID-19 pandemic. Identifying a multi-target-directed ligand approach would open up new opportunities for drug discovery to combat COVID-19. The aim of this work was to perform a virtual screening of an exclusive chemical library of about 1700 molecules containing both pharmacologically active compounds and synthetic intermediates to propose potential protein inhibitors for use against SARS-CoV-2. In silico analysis showed that our compounds triggered an interaction network with key residues of the SARS-CoV-2 spike protein (S-protein), blocking trimer formation and interaction with the human receptor hACE2, as well as with the main 3C-like protease (3CLpro), inhibiting their biological function. Our data may represent a step forward in the search for potential new chemotherapeutic agents for the treatment of COVID-19.

As the incidence of pancreatic ductal adenocarcinoma (PDAC) continues to grow, so does the need for new strategies for treatment. One such area being evaluated is cryoablation. While promising, studies remain limited and questions surrounding basic dosing (minimal lethal temperature) coupled with technological issues associated with accessing PDAC tumors and tumor proximity to vasculature and bile ducts, among others, have limited the use of cryoablation. Additionally, as chemotherapy remains the first-line of attack for PDAC, there is limited information on the impact of combining freezing with chemotherapy. As such, this study investigated the in vitro response of a PDAC cell line to freezing, chemotherapy, and the combination of chemotherapy pre-treatment and freezing. PANC-1 cells and PANC-1 tumor models were exposed to cryoablation (freezing insult) and compared to non-frozen controls. Additionally, PANC-1 cells were exposed to varying sub-clinical doses of gemcitabine or oxaliplatin alone and in combination with freezing. The results show that freezing to −10 °C did not affect viability, whereas −15 °C and −20 °C resulted in a reduction in 1 day post-freeze viability to 85% and 20%, respectively, though both recovered to controls by day 7. A complete cell loss was found following a single freeze below −25 °C. The combination of 100 nM gemcitabine (1.1 mg/m2) pre-treatment and a single freeze at −15 °C resulted in near-complete cell death (2) pre-treatment and a single −15 °C freeze resulted in a similar trend of increased PANC-1 cell death. In summary, these in vitro results suggest that freezing alone to temperatures in the range of −25 °C results in a high degree of PDAC destruction. Further, the data support a potential combinatorial chemo/cryo-therapeutic strategy for the treatment of PDAC. These results suggest that a reduction in chemotherapeutic dose may be possible when offered in combination with freezing for the treatment of PDAC.

Climate change threats make it difficult to perform reliable and quick predictions on floods forecasting. This gives rise to the need of having advanced methods, e.g., computational intelligence tools, to improve upon the results from flooding events simulations and, in turn, design best practices for riverbed maintenance. In this context, being able to accurately estimate the roughness coefficient, also known as Manning’s n coefficient, plays an important role when computational models are employed. In this piece of research, we propose an optimal approach for the estimation of ‘n’. First, an objective function is designed for measuring the quality of ‘candidate’ Manning’s coefficients relative to specif cross-sections of a river. Second, such function is optimised to return coefficients having the highest quality as possible. Five well-known meta-heuristic algorithms are employed to achieve this goal, these being a classic Evolution Strategy, a Differential Evolution algorithm, the popular Covariance Matrix Adaptation Evolution Strategy, a classic Particle Swarm Optimisation and a Bayesian Optimisation framework. We report results on two real-world case studies based on the Italian rivers ‘Paglia’ and ‘Aniene’. A comparative analysis between the employed optimisation algorithms is performed and discussed both empirically and statistically. From the hydrodynamic point of view, the experimental results are satisfactory and produced within significantly less computational time in comparison to classic methods. This shows the suitability of the proposed approach for optimal estimation of the roughness coefficient and, in turn, for designing optimised hydrological models.

Surgical site infection (SSI) substantially contributes each year to patients’ morbidity and mortality, accounting for about 15% of all nosocomial infections. SSI drastically increases the rehab stint and expenses while jeopardizing health outcomes. Besides prevention, the treatment regime relies on an adequate antibiotic therapy. On the other hand, resistant bacterial strains have currently reached up to 34.3% of the total infections, and this percentage grows annually, reducing the efficacy of the common treatment schemes. Thus, new antibacterial strategies are urgently demanded. Here, we demonstrated in rats the effectiveness of non-persistent silver nano-architectures (AgNAs) in infected wound healing together with their synergistic action in combination with chlorhexidine. Besides the in vivo efficacy evaluation, we performed analysis of the bacteriological profile of purulent wound, histological evaluations, and macrophages polarization quantifications to further validate our findings and elucidate the possible mechanisms of AgNAs action on wound healing. These findings open the way for the composition of robust multifunctional nanoplatforms for the translation of safe and efficient topical treatments of SSI.

Increasing frequency of native jellyfish proliferations and massive appearance of non-indigenous jellyfish species recently concur to impact Mediterranean coastal ecosystems and human activities at sea. Nonetheless, jellyfish biomass may represent an exploitable novel resource to coastal communities, with reference to its potential use in the pharmaceutical, nutritional, and nutraceutical Blue Growth sectors. The zooxanthellate jellyfish Cassiopea andromeda, Forsskål, 1775 (Cnidaria, Rhizostomeae) entered the Levant Sea through the Suez Canal and spread towards the Western Mediterranean to reach Malta, Tunisia, and recently also the Italian coasts. Here we report on the biochemical characterization and antioxidant activity of C. andromeda specimens with a discussion on their relative biological activities. The biochemical characterization of the aqueous (PBS) and hydroalcoholic (80% ethanol) soluble components of C. andromeda were performed for whole jellyfish, as well as separately for umbrella and oral arms. The insoluble components were hydrolyzed by sequential enzymatic digestion with pepsin and collagenase. The composition and antioxidant activity of the insoluble and enzymatically digestible fractions were not affected by the pre-extraction types, resulting into collagen- and non-collagen-derived peptides with antioxidant activity. Both soluble compounds and hydrolyzed fractions were characterized for the content of proteins, phenolic compounds, and lipids. The presence of compounds coming from the endosymbiont zooxanthellae was also detected. The notable yield and the considerable antioxidant activity detected make this species worthy of further study for its potential biotechnological sustainable exploitation.

This work introduces an automatic classification system for measuring the complexity level of a given Italian text under a linguistic point-of-view. The task of measuring the complexity of a text is cast to a supervised classification problem by exploiting a dataset of texts purposely produced by linguistic experts for second language teaching and assessment purposes. The commonly adopted Common European Framework of Reference for Languages (CEFR) levels were used as target classification classes, texts were elaborated by considering a large set of numeric linguistic features, and an experimental comparison among ten widely used machine learning models was conducted. The results show that the proposed approach is able to obtain a good prediction accuracy, while a further analysis was conducted in order to identify the categories of features that influenced the predictions.

Pleural empyema represents a severe complication of community acquired pneumonia with an incidence of 0.6% among hospitalized children. Clinical manifestations of picture may be different in infants and young children and it should always be suspected in a child with pneumonia without significant clinical improvement after 48 hours of antibiotic treatment. The most common microorganism associated with empyema is Streptococcus pneumoniae, especially in children under 5 years of age. Chest radiograph is the gold standard for diagnosis but chest ultrasonography, and in some cases chestCT, may be necessary to study features and evolution of the pleural fluid in order to guide therapeutic choices. In most cases small pleural empyema responds to antibiotics alone. However in severe and extensive cases, drainage and invasive treatments, like video-assisted thoracic surgery (VATS) and thoracotomy, became necessary. Due to the early start of antibiotic therapy, blood and pleural fluid cultures may result negative; in these cases only the use of molecular techniques, like polymerase chain reaction in biological fluids, may determine the etiology of the infection. Here we report the case of previously healthy 8-years-old boy with an important and severe pleural empyema as a complication of S. Pneumoniae pneumonia, that did not respond to antibiotic therapy and thoracocentesis and for which decortication has been necessary. In our patient only molecular analysis on pleuric fluid has allowed us to define the etiology of the process.

Chronic granulomatous disease is a rare, inherited immunodeficiency caused by deletions or mutations in genes that encode subunits of the NADPH oxidase complex. The pattern of chronic granulomatous disease inheritance can be X-linked (about 70% of cases) or autosomal recessive. The basic defect lies in phagocytic cells (neutrophils and monocytes) which fail to effectively destroy invading bacteria and fungi. Also, a dysregulated immune response, characterized by extensive granulomatous inflammation of visceral organs, develops in patients. This immunodeficiency is characterized by repeated suppurative infections mainly located in the lungs, skin, and lymph nodes, but also affecting other organs. The major agents involved in the infections are catalase positive bacteria, mycobacteria, fungi, and other opportunistic germs. Diagnosis is based on clinical suspicion and confirmed by nitroblue tetrazolium test or flow cytometry that demonstrate the inability of phagocytes from affected individuals to produce superoxides. The treatment of chronic granulomatous disease involves, in addition to general care such as the prevention of infections and vaccinations, the use of sulfamethoxazole–trimethoprim in combination with itraconazole for prophylaxis. We report the case of a 3-year-old boy with medical history of recurrent respiratory infections, anemia, growth failure, elevated inflammatory markers and occasional rectal bleeding. He was admitted to our department for a suspected chronic bowel inflammatory disease. Clinical history, lymph nodes involvement and the discovery of intestinal granulomas on biopsies confirmed the diagnosis of chronic granulomatous disease.

The occurrence of dysphagia in a child may be a sign of various pathological conditions and mainly gastrointestinal disorders; neurological causes are not very frequent, but they should be taken in to considerations. Etiological diagnosis is important for minimizing related complications. Here we report the case of a 6-year-old girl who was admitted to our Clinic with sudden onset weakness of the limbs and dysphagia. Physical examination revealed hypo-areflexia of both legs and arms and multiple cranial nerve dysfunction. Based on typical clinical course, laboratory investigations and electrophysiological studies, a diagnosis of Guillain-Barrè Syndrome (GBS) was assessed. A treatment with intravenous immunoglobulin (IVIG) was immediately started with a progressive recovery of motility and cranial nerve function. An electrophysiological evaluation, performed one month after therapy start, showed slight improvement of neurological symptoms, in particular of the sensitive component. On the basis of our experience we suggest that a GBS should be suspected when dysphagia is associated with pain and ascending flaccid paralysis of the limbs, in order to prevent a severe complications such as respiratory failure.

Congenital Adrenal hypoplasia (CAH) is a rare genetic disorder which can present two distinct modalities of transmission: recessive X-linked or recessive autosomal modality. That linked to the X chromosome is generally associated with hypogonadotropic hypogonadism (HH). In this case, the gene responsible is DAX-1. We report the history of an infant 36-days old who was admitted to our clinic complaining of acute adrenal insufficiency (AAI), salt wasting and metabolic acidosis. He was the nephew of one of our patients with X-linked CAH. The main causes of early onset AAI were excluded and on the basis of family history we suspected diagnosis of X-linked CAH. Therefore, replacement therapy with gluco- and mineral-corticoids was started immediately. Molecular analyses confirmed the occurrence of DAX-1 gene mutation F449fsX461. His maternal uncle was already recognized to suffer from this disorder, while his mother, grandmother and maternal aunt carried the DAX-1 mutation in heterozygosis. This case report stress the role of genetic counseling in families with DAX-1 gene mutation

Marine algae have gained much importance in the development of nutraceutical products due to their high content of bioactive compounds. In this work, we investigated the activity of Padina pavonica with the aim to demonstrate the pro-osteogenic ability of its extract on human primary osteoblast (HOb). Our data indicated that the acetonic extract of P. pavonica (EPP) is a safe product as it did not show any effect on osteoblast viability. At the same time, EPP showed to possess a beneficial effect on HOb functionality, triggering their differentiation and mineralization abilities. In particular EPP enhanced the expression of the earlier differentiation stage markers: a 5.4-fold increase in collagen type I alpha 1 chain (COL1A1), and a 2.3-fold increase in alkaline phosphatase (ALPL), as well as those involved in the late differentiation stage: a 3.7-fold increase in osteocalcin (BGLAP) expression and a 2.8-fold in osteoprotegerin (TNFRSF11B). These findings were corroborated by the enhancement in ALPL enzymatic activity (1.7-fold increase) and by the reduction of receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) ratio (0.6-fold decrease). Moreover, EPP demonstrated the capacity to enhance the bone nodules formation by 3.2-fold in 4 weeks treated HOb. Therefore, EPP showed a significant capability of promoting osteoblast phenotype. Given its positive effect on bone homeostasis, EPP could be used as a useful nutraceutical product that, in addition to a healthy lifestyle and diet, can be able to contrast and prevent bone diseases, especially those connected with ageing, such as osteoporosis (OP).

Recently, seaweeds and their extracts have attracted great interest in the pharmaceutical industry as a source of bioactive compounds. Studies have demonstrated the cytotoxic activity of macroalgae towards different types of cancer cell models, and their consumption has been suggested as a chemo-preventive agent against several cancers such as breast, cervix and colon cancers. Reports relevant to the chemical properties of brown algae Padina sp. are limited and those accompanied to a comprehensive evaluation of the biological activity on osteosarcoma (OS) are non existent. In this report, we explored the chemical composition of French Polynesian Padina pavonica extract (EPP) by spectrophotometric assays (total phenolic, flavonoid and tannin content, and antioxidant activity) and by gas chromatography-mass spectrometry (GC-MS) analysis, and provided EPP lipid and sterols profiles. Several compounds with relevant biological activity were also identified that suggest interesting pharmacological and health-protecting effects for EPP. Moreover, we demonstrated that EPP presents good anti-proliferative and pro-apoptotic activities against two OS cell lines, SaOS-2 and MNNG, with different cancer-related phenotypes. Finally, our data suggest that EPP might target different properties associated with cancer development and aggressiveness.

N-Methyl-D-aspartic acid (NMDA) receptors are widely expressed in the brain and are critical for many forms of synaptic plasticity. Subtypes of the NMDA receptor NR2 subunit are differentially expressed during development; in the forebrain, the NR2B receptor is dominant early in development, and later both NR2A and NR2B are expressed. In heterologous expression systems, NR2A-containing receptors open more reliably and show much faster opening and closing kinetics than do NR2B-containing receptors. However, conflicting data, showing similar open probabilities, exist for receptors expressed in neurons. Similarly, studies of synaptic plasticity have produced divergent results, with some showing that only NR2A-containing receptors can drive long-term potentiation and others showing that either subtype is capable of driving potentiation. In order to address these conflicting results as well as open questions about the number and location of functional receptors in the synapse, we constructed a Monte Carlo model of glutamate release, diffusion, and binding to NMDA receptors and of receptor opening and closing as well as a model of the activation of calcium-calmodulin kinase II, an enzyme critical for induction of synaptic plasticity, by NMDA receptor-mediated calcium influx. Our results suggest that the conflicting data concerning receptor open probabilities can be resolved, with NR2A- and NR2B-containing receptors having very different opening probabilities. They also support the conclusion that receptors containing either subtype can drive long-term potentiation. We also are able to estimate the number of functional receptors at a synapse from experimental data. Finally, in our models, the opening of NR2B-containing receptors is highly dependent on the location of the receptor relative to the site of glutamate release whereas the opening of NR2A-containing receptors is not. These results help to clarify the previous findings and suggest future experiments to address open questions concerning NMDA receptor function.

Reaching and grasping in primates depend on the coordination of neural activity in large frontoparietal ensembles. Here we demonstrate that primates can learn to reach and grasp virtual objects by controlling a robot arm through a closed-loop brain-machine interface (BMIc) that uses multiple mathematical models to extract several motor parameters (i.e., hand position, velocity, gripping force, and the EMGs of multiple arm muscles) from the electrical activity of frontoparietal neuronal ensembles. As single neurons typically contribute to the encoding of several motor parameters, we observed that high BMIc accuracy required recording from large neuronal ensembles. Continuous BMIc operation by monkeys led to significant improvements in both model predictions and behavioral performance. Using visual feedback, monkeys succeeded in producing robot reach-and-grasp movements even when their arms did not move. Learning to operate the BMIc was paralleled by functional reorganization in multiple cortical areas, suggesting that the dynamic properties of the BMIc were incorporated into motor and sensory cortical representations.