Antibody responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are multi-targeted and variable over time. Multiplex quantitative serological assays are needed to provide accurate and robust seropositivity data for the establishment of serological signatures during vaccination and or infection. We describe here the validation and evaluation of an electro-chemiluminescence (ECL)-based Mesoscale Discovery assay (MSD) for estimation of total and functional IgG relative to SARS-CoV-2 spike, nucleocapsid and receptor binding (RBD) proteins in human serum samples to establish serological signatures of SARS-CoV-2 natural infection and breakthrough cases. The 9-PLEX assay was validated as per ICH, EMA, and US FDA guidelines using a panel of sera samples, including the NIBSC/WHO reference panel (20/268). The assay demonstrated high specificity and selectivity in inhibition assays, wherein the homologous inhibition was more than 85% and heterologous inhibition was below 10%. The assay also met predetermined acceptance criteria for precision (CV < 20%), accuracy (70–130%) and dilutional linearity. The method’s applicability to serological signatures was demonstrated using sera samples (n = 45) representing vaccinated, infected and breakthrough cases. The method was able to establish distinct serological signatures and thus provide a potential tool for seroprevalence of SARS-CoV-2 during vaccination or infection. Full article

While typically exhibiting characteristic features, fungal infections can sometimes present in an unusual context, having improbable localization (eyelid, face, or joint); mimicking other skin diseases such as eczema, psoriasis, or mycosis fungoides; and appearing with unexpected color, shape, or distribution. The emergence of such a challenging clinical picture is attributed to the complex interplay of host characteristics (hygiene and aging population), environment (climate change), advances in medical procedures, and agent factors (fungal resistance and species emergence). We aim to provide a better understanding of unusual epidemiological contexts and atypical manifestations of fungal superficial diseases, knowing that there is no pre-established clinical guide for these conditions. Thus, a literature examination was performed to provide a comprehensive analysis on rare and atypical superficial mycosis as well as an update on certain fungal clinical manifestations and their significance. The research and standard data extraction were performed using PubMed, Medline, Scopus, and EMBASE databases, and a total of 222 articles were identified. This review covers published research findings for the past six months. Full article

The burgeoning complexity and heterogeneity of IoT networks, coupled with their rapid growth, constant evolution, and new players, present significant challenges in terms of connectivity, interoperability, management, and usability. These networks, composed of a diverse array of devices, technologies and the like, demand innovative solutions to bridge the gaps between different IoT technologies and communication protocols. This article presents a simple, yet efficacious communication Relay to address one of these critical gaps. This Relay uses NB-IoT to ease the integration of 6LoWPAN-based IoT devices (IPv6) into the public legacy Internet (IPv4). This device translates 6LoWPAN, IPv6 CoAP messages into Internet-standard REST requests, so that appropriate handling of devices’ data be achieved in several stages. Thus, the Relay establishes two branches of communications: (i) the local network where the 6LoWPAN gateway is placed, and (ii) the public NB-IoT network. User interaction and data analysis are achieved by virtue of Home Assistant, where former 6LoWPAN devices are now discovered and shown as proper Home Assistant entities thanks to the Relay’s ease of integration into the open-source platform. This novel approach not only ensures efficient data and network management, but it also meets the urgent necessity for advanced solutions in enhancing actual IoT interconnectivity and monitoring. The unprecedented pace at which IoT devices, players and different networks have been proliferating in recent times is not compatible with countless manufacturer-dependent platforms, applications, and proprietary protocols that the IoT field has been leading with so far, almost from its beginnings. Full article

Decades of research has revealed a relationship between childhood socioeconomic status (SES) and brain development at the structural and functional levels. Of particular note is the distinction between income and maternal education, two highly correlated factors which seem to influence brain development through distinct pathways. Specifically, while a families’ income-to-needs ratio is linked with physiological stress and household chaos, caregiver education influences the day-to-day language environment a child is exposed to. Variability in either one of these environmental experiences is related to subsequent brain development. While this work has the potential to inform public policies in a way that benefits children, it can also oversimplify complex factors, unjustly blame low-SES parents, and perpetuate a harmful deficit perspective. To counteract these shortcomings, researchers must consider sociodemographic differences in the broader cultural context that underlie SES-based differences in brain development. This review aims to address these issues by (a) identifying how sociodemographic mechanisms associated with SES influence the day-to-day experiences of children, in turn, impacting brain development, while (b) considering the broader cultural contexts that may differentially impact this relationship. Full article

In this study, composite particles of rGO/CoFe₂O₄ were synthesized using a solvothermal method to fabricate a low-density magnetorheological (MR) material with enhanced sedimentation stability. The morphology and crystallographic features of rGO/CoFe₂O₄ were characterized via SEM, TEM, and XRD, and its magnetic properties were tested using VSM. The MR fluid was formulated by blending rGO/CoFe₂O₄ particles into silicone oil. Under different magnet strengths (H), a rotational rheometer was used to test its MR properties. Typical MR properties were observed, including shear stress, viscosity, storage/loss modulus, and dynamic yield stress (${\tau }_{dy}$) following the Herschel–Bulkley model reaching 200 Pa when H is 342 kA/m. Furthermore, the yield stress of the MR fluid follows a power law relation as H increases and the index changes from 2.0 (in the low H region) to 1.5 (in the high H region). Finally, its MR efficiency was calculated to be about 10⁴% at H of 342 kA/m. Full article

COVID-19 is a lung disease caused by a coronavirus family virus. Due to its extraordinary prevalence and associated death rates, it has spread quickly to every country in the world. Thus, achieving peaks and outlines and curing different types of relapses is extremely important. Given the worldwide prevalence of coronavirus and the participation of physicians in all countries, information has been gathered regarding the properties of the virus, its diverse types, and the means of analyzing it. Numerous approaches have been used to identify this evolving virus. It is generally considered the most accurate and acceptable method of examining the patient’s lungs and chest through a CT scan. As part of the feature extraction process, a method known as fractional Fourier transform (FrFT) has been applied as one of the time-frequency domain transformations. The proposed method was applied to a database consisting of 2481 CT images. Following the transformation of all images into equal sizes and the removal of non-lung areas, multiple combination windows are used to reduce the number of features extracted from the images. In this paper, the results obtained for KNN and SVM classification have been obtained with accuracy values of 99.84% and 99.90%, respectively. Full article

In recent years, the incidence of fungal infections in humans has increased dramatically, accompanied by an expansion in the number of species implicated as etiological agents, especially environmental fungi never involved before in human infection. Among fungal pathogens, Candida species are the most common opportunistic fungi that can cause local and systemic infections, especially in immunocompromised individuals. Candida albicans (C. albicans) is the most common causative agent of mucosal and healthcare-associated systemic infections. However, during recent decades, there has been a worrying increase in the number of emerging multi-drug-resistant non-albicans Candida (NAC) species, i.e., C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. auris, and C. ciferrii. In particular, Candida ciferrii, also known as Stephanoascus ciferrii or Trichomonascus ciferrii, is a heterothallic ascomycete yeast-like fungus that has received attention in recent decades as a cause of local and systemic fungal diseases. Today, the new definition of the S. ciferrii complex, which consists of S. ciferrii, Candida allociferrii, and Candida mucifera, was proposed after sequencing the 18S rRNA gene. Currently, the S. ciferrii complex is mostly associated with non-severe ear and eye infections, although a few cases of severe candidemia have been reported in immunocompromised individuals. Low susceptibility to currently available antifungal drugs is a rising concern, especially in NAC species. In this regard, a high rate of resistance to azoles and more recently also to echinocandins has emerged in the S. ciferrii complex. This review focuses on epidemiological, biological, and clinical aspects of the S. ciferrii complex, including its pathogenicity and drug resistance. Full article

The tumor-to-stroma ratio is a highly debated prognostic factor in the management of several solid tumors and there is no universal agreement on its practicality. In our study, we proposed confirming or dismissing the hypothesis that a simple measurement of stroma quantity is an easy-to-use and strong prognostic tool. We have included 74 consecutive patients with colorectal cancer who underwent primary curative abdominal surgery. The tumors have been grouped into stroma-poor (stroma < 10%), medium-stroma (between 10 and 50%) and stroma-rich (over 50%). The proportion of tumor stroma ranged from 5% to 70% with a median of 25%. Very few, only 6.8% of patients, had stroma-rich tumors, 4% had stroma-poor tumors and 89.2% had tumors with a medium quantity of stroma. The proportion of stroma, at any cut-off, had no statistically significant influence on the disease-specific survival. This can be explained by the low proportion of stroma-rich tumors in our patient group and the inverse correlation between stroma proportion and tumor grade. The real-life proportion of stroma-rich tumors and the complex nature of the stroma–tumor interaction has to be further elucidated. Full article

Northeast China’s sandy region is an arid and semi-arid zone highly susceptible to climate change. Investigating the long-term changes in the Northeast China sandy land (Northeast China sandy land, DBSL) landscape can provide an important basis for the ecological restoration of this region. This study analyzed long-term remote sensing data of the DBSL from 1980 to 2022 and explored the spatial pattern, evolution, and driving mechanisms. In 2022, vegetation was mainly distributed in the northwest, center, and southwest, covering a total area of 30,508.82 km². Areas with high and medium vegetation cover showed strong aggregation characteristics and were mainly distributed in the southwest, whereas those with low vegetation coverage were highly dispersed and widely distributed in the central region. Lakes were widely distributed in the northwest and central regions, with a total area of 2736.43 km². In the last 42 years, the vegetation cover decreased by 24.48%. Areas with high and medium vegetation coverage decreased in size, and those with low vegetation coverage first increased and then decreased, with overall decreases of 35.35%, 19.16%, and 6.88%, respectively. The overall area of the DBSL showed various degrees of degradation. Shrinking and dry lakes were concentrated in the sandy hinterland. The lake landscape changed significantly from 1990 to 2010, with a decrease in lake area of 27.41%. In contrast, the sandy area increased by 25.65%, indicating a high degree of desertification. However, from 2005 to 2022, desertification decelerated. The most important factors driving the evolution of the DBSL were socio-economic factors. The increase in human disturbance will have a certain impact on the landscape changes in the region in the short term. The national policy of returning farmland to fields and grasslands will affect the increase of vegetation and lake landscape area in the short term, and the sand area and excessive animal husbandry will be reduced. This study provides a scientific basis for ecological restoration and sustainable development in Northeast China. Full article

Context: Psychological distress symptoms in the last days of life often contribute to the overall symptom burden in frail older patients. Good symptom management practices are crucial to ensure high-quality end-of-life care in an aging population, though the best pharmacological approach to treat these psychological symptoms has yet to be established. Objectives: To identify current evidence-based and practice-based knowledge of pharmacological interventions for the treatment of agitation, delirium, and anxiety during the last days of life in frail older patients. Methods: A systematic, mixed methods review was performed through MEDLINE via PubMed and EMBASE from inception until February 2022 and updated through March 2023. National and international guideline databases and grey literature were searched for additional studies and guidelines. Results: Four quantitative studies, two non-randomized and two descriptive, were identified. No randomized controlled trials met inclusion criteria. No qualitative studies were withheld. The three consensus-based protocols that were found through citation searching and screening of grey literature did not meet the standards for inclusion. Haloperidol is recommended in consensus-based guidelines for delirium and is widely used, but high-quality evidence about its efficacy is missing. Better control of agitation or refractory delirium might be achieved with the addition of a benzodiazepine. There is no evidence available about the treatment of anxiety in the last days of life in frail older patients. Conclusions: This mixed methods review demonstrates the lack of good quality evidence that is needed to help clinicians with pharmacological treatment decisions when confronted with psychological symptoms in the last days of life in frail older patients. Population aging will only emphasize the need for further research in this specific population. Full article

Liver cancer is the second leading cause of cancer-related death in males. It is estimated that approximately one million deaths will occur by 2030 due to hepatic cancer. Hepatocellular carcinoma (HCC) is the most prevalent primary liver cancer subtype and is commonly diagnosed at an advanced stage. The drug arsenal used in systemic therapy for HCC is very limited. Multikinase inhibitors sorafenib (Nexavar^®) and lenvatinib (Lenvima^®) have been used as first-line drugs with modest therapeutic effects. In this scenario, it is imperative to search for new therapeutic strategies for HCC. Herein, the antiproliferative activity of N-acylhydrazone derivatives was evaluated on HCC cells (HepG2 and Hep3B), which were chemically planned on the ALL-993 scaffold, a potent inhibitor of vascular endothelial growth factor 2 (VEGFR2). The substances efficiently reduced the viability of HCC cells, and the LASSBio-2052 derivative was the most effective. Further, we demonstrated that LASSBio-2052 treatment induced FOXM1 downregulation, which compromises the transcriptional activation of genes required for G2/M transition, such as AURKA and AURKB, PLK1, and CDK1. In addition, LASSBio-2052 significantly reduced CCNB1 and CCND1 expression in HCC cells. Our findings indicate that LASSBio-2052 is a promising prototype for further in vivo studies. Full article

The importance of spirituality in the treatment of mental illness is increasingly acknowledged, but mental healthcare professionals often feel they lack specific competence. An instrument is missing to quantify the spiritual care competence of mental healthcare professionals in the Netherlands. The aim of this study was thus to translate the Spiritual Care Competence Questionnaire (SCCQ) into Dutch and validate it for use in mental healthcare. After translation, the SCCQ-NL was distributed in a cross-sectional design among 3497 healthcare professionals in two mental healthcare institutions (MHIs) in the Netherlands. In the sample of 730 completed questionnaires, exploratory factor analysis revealed seven factors: perception of spiritual needs competencies, team spirit, spiritual self-awareness, documentation competencies, empowerment and proactive opening competencies, knowledge about other religions, and conversation competencies. One item was deleted during the process. Internal consistency for the 25-item SCCQ-NL subscales is sufficient with Cronbach’s alpha ranging from 0.64 to 0.81. Conversation competencies and perception of spiritual needs scored highest in the sample, next to knowledge about other religions and empowerment competencies, while spiritual self-awareness, team spirit and documentation competencies scored the lowest. Small but significant differences in several subscale scores were found for profession, identifying oneself as a believer, practicing prayer and/or meditation, age and working years. The SCCQ-NL can be used for the assessment of spiritual care competencies and for the planning and evaluation of training and improvement strategies. Full article

Beaches are among the habitats most frequented by migratory birds for breeding and/or wintering. However, threats such as human pressure and sea level rise can reduce the availability of these habitats for different species. The presence of alternative areas, such as salt pans and brackish habitats, is essential for many migratory shorebird populations. This study addresses the post-breeding dispersal of the Kentish plover (Charadrius alexandrinus) in the Iberian Peninsula by analysing C and N isotopes in feathers. The study was conducted at six locations along the Iberian coast, which were categorized into three areas: the NW Atlantic coast, the Atlantic coast of Andalusia, and the Mediterranean coast. Although linear mixed models did not reveal any significant effects of sex or coastal area on isotopic levels, the variability in the data suggests different habitat-use strategies in the post-reproductive period. Isotopic levels in birds from the northwest of the Iberian Peninsula exhibit greater fidelity to a single habitat type, while those from the Mediterranean coast and the Atlantic coast of Andalusia show greater variability, indicating different individual dispersal strategies. The lack of alternative habitats for the northwest Iberian population, the reduction in available habitat due to rising sea levels, and human pressure together pose a serious threat to the survival of this species, already with an unfavourable conservation status. Full article

A minimalist approach to the linear stability problem in fluid dynamics is developed that ensures efficiency by utilizing only the essential elements required to find the eigenvalues for given boundary conditions. It is shown that the problem is equivalent to a single first-order ordinary differential equation, and that studying the argument of the unknown complex function in the eigenvalue space is sufficient to find the dispersion relation. The method is applied to a model for relativistic magnetized astrophysical jets. Full article

This review examines the impact of obesity on the pathophysiology of heart failure with preserved ejection fraction (HFpEF) and focuses on novel mechanisms for HFpEF prevention using a glucagon-like peptide-1 receptor agonism (GLP-1 RA). Obesity can lead to HFpEF through various mechanisms, including low-grade systemic inflammation, adipocyte dysfunction, accumulation of visceral adipose tissue, and increased pericardial/epicardial adipose tissue (contributing to an increase in myocardial fat content and interstitial fibrosis). Glucagon-like peptide 1 (GLP-1) is an incretin hormone that is released from the enteroendocrine L-cells in the gut. GLP-1 reduces blood glucose levels by stimulating insulin synthesis, suppressing islet α-cell function, and promoting the proliferation and differentiation of β-cells. GLP-1 regulates gastric emptying and appetite, and GLP-1 RA is currently indicated for treating type 2 diabetes (T2D), obesity, and metabolic syndrome (MS). Recent evidence indicates that GLP-1 RA may play a significant role in preventing HFpEF in patients with obesity, MS, or obese T2D. This effect may be due to activating cardioprotective mechanisms (the endogenous counter-regulatory renin angiotensin system and the AMPK/mTOR pathway) and by inhibiting deleterious remodeling mechanisms (the PKA/RhoA/ROCK pathway, aldosterone levels, and microinflammation). However, there is still a need for further research to validate the impact of these mechanisms on humans. Full article

Patients with Solid Organ Transplantations (SOTx) face long-term lifestyle adaptations, psychological and social adjustments, and complex self-care regimes to maintain health post-transplant. Self-management (SM) skills represent important aspects of nursing communication with SOTx patients; however, there is potential for SM to be defined narrowly in terms of medication adherence. The study presented here collated the existing definitions in a mixed method review in order to identify SM attributes for this group (including those unique to this population). Secondary analysis of a dataset and bibliographic analysis and an expert panel were used to develop a comprehensive working definition of SOTx patients. The analysis comprised critical interpretation of the evolving definition content, concepts, and contexts of application in current usages and over time. We identified eight definitions and 63 cited definition sources from bibliographic analysis. Findings identified limitations of the existing definitions. Population-specific attributes included optimisation of transplant outcomes, active engagement in healthy behaviours, control, structure, and discipline characteristics, and moderating factors of patient motivation, self-efficacy, and cognitive function. A critical appraisal of definitions indicated inadequately defined aspects such as setting, temporal dimension, concept interaction, interventions, and measurable outcomes. The bibliographic analysis highlighted the influence of broader chronic illness constructions of SM, underpinning the generalisable SM attributes in current definitions. Further research may advance the development of a definition in exploring the relevance of SOTx-specific attributes of the definition. Full article

In the proposed paper, an artificial neural network (ANN) algorithm is applied to predict the electronic circuit outputs of voltage signals in Industry 4.0/5.0 scenarios. This approach is suitable to predict possible uncorrected behavior of control circuits affected by unknown noises, and to reproduce a testbed method simulating the noise effect influencing the amplification of an input sinusoidal voltage signal, which is a basic and fundamental signal for controlled manufacturing systems. The performed simulations take into account different noise signals changing their time-domain trend and frequency behavior to prove the possibility of predicting voltage outputs when complex signals are considered at the control circuit input, including additive disturbs and noises. The results highlight that it is possible to construct a good ANN training model by processing only the registered voltage output signals without considering the noise profile (which is typically unknown). The proposed model behaves as an electronic black box for Industry 5.0 manufacturing processes automating circuit and machine tuning procedures. By analyzing state-of-the-art ANNs, the study offers an innovative ANN-based versatile solution that is able to process various noise profiles without requiring prior knowledge of the noise characteristics. Full article

We propose a new approach for volumetric instance segmentation in X-ray Computed Tomography (CT) data for Non-Destructive Testing (NDT) by combining the Segment Anything Model (SAM) with tile-based Flood Filling Networks (FFN). Our work evaluates the performance of SAM on volumetric NDT data-sets and demonstrates its effectiveness to segment instances in challenging imaging scenarios. We implemented and evaluated techniques to extend the image-based SAM algorithm for the use with volumetric data-sets, enabling the segmentation of three-dimensional objects using FFN’s spatial adaptability. The tile-based approach for SAM leverages FFN’s capabilities to segment objects of any size. We also explore the use of dense prompts to guide SAM in combining segmented tiles for improved segmentation accuracy. Our research indicates the potential of combining SAM with FFN for volumetric instance segmentation tasks, particularly in NDT scenarios and segmenting large entities and objects. While acknowledging remaining limitations, our study provides insights and establishes a foundation for advancements in instance segmentation in NDT scenarios. Full article

Oxidative phosphorylation involves a complex multi-enzymatic mitochondrial machinery critical for proper functioning of the cell, and defects herein cause a wide range of diseases called “primary mitochondrial disorders” (PMDs). Mutations in about 400 nuclear and 37 mitochondrial genes have been documented to cause PMDs, which have an estimated birth prevalence of 1:5000. Here, we describe a 4-year-old female presenting from early childhood with psychomotor delay and white matter signal changes affecting several brain regions, including the brainstem, in addition to lactic and phytanic acidosis, compatible with Leigh syndrome, a genetically heterogeneous subgroup of PMDs. Whole genome sequencing of the family trio identified a homozygous 12.9 Kb deletion, entirely overlapping the NDUFA4 gene. Sanger sequencing of the breakpoints revealed that the genomic rearrangement was likely triggered by Alu elements flanking the gene. NDUFA4 encodes for a subunit of the respiratory chain Complex IV, whose activity was significantly reduced in the patient’s fibroblasts. In one family, dysfunction of NDUFA4 was previously documented as causing mitochondrial Complex IV deficiency nuclear type 21 (MC4DN21, OMIM 619065), a relatively mild form of Leigh syndrome. Our finding confirms the loss of NDUFA4 function as an ultra-rare cause of Complex IV defect, clinically presenting as Leigh syndrome. Full article

Apple cultivar classification is challenging due to the inter-class similarity and high intra-class variations. Human experts do not rely on single-view features but rather study each viewpoint of the apple to identify a cultivar, paying close attention to various details. Following our previous work, we try to establish a similar multiview approach for machine-learning (ML)-based apple classification in this paper. In our previous work, we studied apple classification using one single view. While these results were promising, it also became clear that one view alone might not contain enough information in the case of many classes or cultivars. Therefore, exploring multiview classification for this task is the next logical step. Multiview classification is nothing new, and we use state-of-the-art approaches as a base. Our goal is to find the best approach for the specific apple classification task and study what is achievable with the given methods towards our future goal of applying this on a mobile device without the need for internet connectivity. In this study, we compare an ensemble model with two cases where we use single networks: one without view specialization trained on all available images without view assignment and one where we combine the separate views into a single image of one specific instance. The two latter options reflect dataset organization and preprocessing to allow the use of smaller models in terms of stored weights and number of operations than an ensemble model. We compare the different approaches based on our custom apple cultivar dataset. The results show that the state-of-the-art ensemble provides the best result. However, using images with combined views shows a decrease in accuracy by 3% while requiring only 60% of the memory for weights. Thus, simpler approaches with enhanced preprocessing can open a trade-off for classification tasks on mobile devices. Full article

Acute respiratory diseases in felines can be attributed to a diverse range of pathogens. The recent emergence of novel viruses, particularly SARS-CoV-2 and its variants, has also been associated with respiratory ailments in cats and other pets, underscoring the need for a highly sensitive diagnostic assay capable of concurrently detecting multiple respiratory pathogens. In this study, we developed a targeted next generation sequencing panel using Ion Torrent Ampliseq technology to detect multiple respiratory pathogens, including recent SARS-CoV-2 variants and Feline herpesvirus-1, Feline calicivirus, Bordetella bronchiseptica, Mycoplasmopsis (previously Mycoplasma) felis, and Chlamydia felis. A PCR amplification-based library preparation, employing primers designed for pathogen target regions, was synthesized and divided into two pools, followed by sequencing and assembly to a repertoire of target pathogen genomes. Analytical sensitivity was assessed based on Ct values from real-time PCR for the corresponding pathogens, indicating an equivalent detection limit. Most of the pathogens under study were positively identified to a limit of approximately Ct 36, whereas for Feline herpesvirus-1 and SARS-CoV-2, positive reads were observed in samples with a Ct of 37. Based on a limited number of samples, the diagnostic sensitivity values for the SARS-CoV-2, Feline herpesvirus-1, and M. felis samples were 100% with no false negative results. The diagnostic specificity of SARS-CoV-2, Feline herpesvirus-1, Feline calicivirus, and C. felis were 100%. Importantly, none of the target primers exhibited non-specific amplification, ensuring the absence of false positive results for other pathogens within the study. Additionally, the assay’s specificity was validated by cross-referencing the raw sequencing data with established databases like BLAST, affirming the high specificity of the targeted Next-Generation Sequencing (tNGS) assay. Variations in the sequencing reads of different pathogens were observed when subjected to diverse extraction methods. Rigorous assessment of the assay’s reliability involved reproducibility across testing personnel and repeated runs. The developed assay’s clinical applicability was tested using samples submitted to the diagnostic laboratory from cat shelters and suspected cases. The developed targeted next-generation sequencing methodology empowers the detection of multiple respiratory pathogens manifesting similar clinical symptoms while offering confirmation of results through genome sequencing. Full article

Osteoarthritis (OA) is a degenerative joint disease, causing impaired mobility. There are currently no effective therapies other than palliative treatment. Mesenchymal stromal cells (MSCs) and their secreted extracellular vesicles (MSC-EVs) have shown promise in attenuating OA progression, promoting chondral regeneration, and modulating joint inflammation. However, the precise molecular mechanism of action driving their beneficial effects has not been fully elucidated. In this study, we analyzed MSC-EV-treated human OA chondrocytes (OACs) to assess viability, proliferation, migration, cytokine and catabolic protein expression, and microRNA and mRNA profiles. We observed that MSC-EV-treated OACs displayed increased metabolic activity, proliferation, and migration compared to the controls. They produced decreased proinflammatory (Il-8 and IFN-γ) and increased anti-inflammatory (IL-13) cytokines, and lower levels of MMP13 protein coupled with reduced expression of MMP13 mRNA, as well as negative microRNA regulators of chondrogenesis (miR-145-5p and miR-21-5p). In 3D models, MSC-EV-treated OACs exhibited enhanced chondrogenesis-promoting features (elevated sGAG, ACAN, and aggrecan). MSC-EV treatment also reversed the pathological impact of IL-1β on chondrogenic gene expression and extracellular matrix component (ECM) production. Finally, MSC-EV-treated OACs demonstrated the enhanced expression of genes associated with cartilage function, collagen biosynthesis, and ECM organization and exhibited a signature of 24 differentially expressed microRNAs, associated with chondrogenesis-associated pathways and ECM interactions. In conclusion, our data provide new insights on the potential mechanism of action of MSC-EVs as a treatment option for early-stage OA, including transcriptomic analysis of MSC-EV-treated OA, which may pave the way for more targeted novel therapeutics. Full article