Search Results (1,154)

Background: Serum-free culture of red blood cells (RBCs) typically leads to rapid loss of viability, limiting experimental and translational applications. Lipid-rich formulations and essential oils may provide biocompatible support for RBC integrity while limiting microbial overgrowth. Methods: RBCs from nine healthy adult donors were cultured in serum-free RPMI under four conditions: control, vehicle (olive oil, 1:100 v/v), genuine adenosine triphosphate (ATP)-oil^® (1:100 v/v), and laboratory oil, “mimicking” ATP-oil^®. Cultures were maintained for 18 days. Viability was assessed by light microscopy and trypan blue exclusion; bacterial contamination was qualitatively observed on day 18. Results: Genuine ATP-oil^® maintained 35–45% RBC viability at day 18, whereas control and vehicle cultures declined rapidly. The mimicking preparation did not reproduce these effects. ATP-oil^® immersion was associated with a qualitative reduction in bacterial contamination versus control, consistent with a dual action on RBC preservation and microbial suppression under serum-free conditions. Conclusions: Supplementation with ATP-oil^® substantially prolongs RBC survival and limits bacterial overgrowth in vitro, outperforming commonly used serum or plasma supplements on a per-volume basis. These findings suggest potential applications for improving ex vivo handling or storage of blood components and for reducing background contamination in diagnostic microbiology. Further studies with larger cohorts are warranted to reveal underlying mechanisms and to define active constituents in order to standardize production. Full article

Rapid detection of β-lactamase activity is becoming increasingly important as β-lactam resistance spreads at an alarming rate and conventional diagnostics often require several hours to deliver actionable results. Over the past decade, methods based on luminescence, bioluminescence, chemiluminescence, and fluorescence have become powerful tools for the functional assessment of resistance resulting from β-lactamase activity. These approaches provide highly sensitive, activity-based readouts, often within minutes, and frequently rely on simple optical instrumentation. In this review, we summarize recent developments in luminescent probe design between 2015 and 2025, with emphasis on reaction mechanisms, analytical performance, and the ability of these systems to discriminate between different β-lactamases, including narrow-spectrum enzymes, AmpC, ESBL, and carbapenemases. We also discuss their applications in bacterial cultures, clinical isolates, complex biological matrices and, in some cases, in vivo models. While luminescent assays are not yet positioned to replace standard susceptibility testing, they offer a practical and increasingly robust complement to culture-based and molecular methods. The emerging trends highlighted here, such as self-immobilizing fluorogenic probes, chemiluminescent relay systems, nanomaterial-based sensors and AI-assisted mobile platforms, suggest that luminescent β-lactamase detection could play a meaningful role in future rapid diagnostics and resistance surveillance. Full article

Fluorescence imaging is crucial for providing detailed information in clinical practice. However, traditional first near-infrared (NIR-I) dyes such as indocyanine green (ICG) exhibit limitations such as shallow penetration depth, low contrast, and suboptimal clarity due to light scattering and autofluorescence. To overcome these drawbacks, we utilized a novel amphiphilic second near-infrared (NIR-II) aggregation-induced emission (AIE) probe (TCP) with an emission range beyond 1300 nm (NIR-IIa). Using approximately 200 co-registered NIR-I/NIR-IIa image pairs acquired with TCP, we trained a SwinUnet-based deep learning model to transform low-quality NIR-I ICG images into high-resolution NIR-IIa-like images. Owing to its superior brightness and photostability, TCP enhances in vivo fluorescent angiography, offering clearer vascular details and a higher signal-to-background ratio (SBR) in the NIR-IIa region, 2.6-fold higher than that of ICG in the NIR-I region. The deep learning model successfully converted blurred NIR-I images into high-SBR NIR-IIa-like images, achieving rapid imaging speeds without compromising quality. This work introduces a synergistic “probe-plus-AI” paradigm that substantially improves both the quality and speed of clinical fluorescence imaging, providing a pathway that is immediately translatable to enhanced diagnostics and image-guided surgery. Full article

Multiphoton endomicroscopy (MPEM) has recently become a key development in optical biomedical diagnostics, providing histologically relevant in vivo images that are eliminating both the need for tissue damage during biopsy sampling and the need for dye injections. Due to its ability to visualize structures at the epithelial, extracellular matrix, and subcellular levels, MPEM offers a promising diagnostic method for precancerous conditions and early forms of gastrointestinal (GI) cancer. The high specificity of multiphoton signals—the two-photon fluorescence response of endogenous fluorophores (NADH, FAD), the second-harmonic generation signal from collagen, and others—makes this method a promising alternative to both traditional histology and confocal endoscopy, enabling real-time assessment of metabolic status, intestinal epithelial cell status, and stromal remodeling. Despite the promising prospects of multiphoton microscopy, its practical implementation is progressing extremely slowly. The main factors here include the difficulty of delivering ultrashort pulses with high peak power, which is necessary for multiphoton excitation (MPE), and obtaining these pulses at the required wavelengths to activate the autofluorescence mechanism. One of the most promising solutions is the use of specialized multimode optical fibers that can both induce beam self-cleaning (BSC), which allows for the formation of a stable beam profile close to the fundamental mode, and significantly broaden the optical spectrum, which can ultimately cover the entire region of interest. This review presents the biophysical foundations of multiphoton microscopy of GI tissue, existing endoscopic architectures for MPE, and an analysis of the potential for using novel nonlinear effects in multimode optical fibers, such as the BSC effect and supercontinuum generation. It is concluded that the use of optical fibers in which the listed effects are realized in the tracts of multiphoton endomicroscopes can become a key step in the creation of a new generation of high-resolution instruments for the early detection of malignant neoplasms of the GI tract. Full article

Tick-borne co-infections are an increasingly recognized and clinically important aspect of Lyme borreliosis, particularly in regions where Ixodes ticks transmit a wide range of bacterial, protozoan, and viral pathogens. In addition to Borrelia burgdorferi sensu lato, these ticks frequently harbor microorganisms such as Babesia spp., Anaplasma phagocytophilum, Ehrlichia spp., Borrelia miyamotoi, Bartonella spp., and several tick-borne viruses. Co-infections may increase disease severity, prolong symptom duration, and contribute to atypical or overlapping clinical presentations, thereby complicating diagnosis and management. Growing evidence from epidemiological studies, clinical case series, and experimental in vivo and in vitro models indicates that pathogen–pathogen and pathogen–host interactions can modulate immune responses and influence disease progression. Diagnostic challenges arise from non-specific clinical features and limitations of current laboratory methods. From a therapeutic perspective, although standard antibiotic regimens for Lyme disease are effective against some bacterial co-infections, they do not provide coverage for protozoan or viral agents, necessitating pathogen-specific and, in some cases, combination treatment strategies. This review synthesizes current knowledge on the epidemiology, clinical impact, diagnostic limitations, and treatment approaches for tick-borne co-infections associated with Lyme disease, and highlights critical evidence gaps and future research directions to improve patient outcomes. Full article

The effective treatment of aggressive brain tumors, such as glioblastoma, is critically hindered by the blood-brain barrier (BBB) and the non-specific clearance of therapeutic agents by the immune system. Superparamagnetic iron oxide nanoparticles (SPMNPs) offer a powerful theranostic platform, combining magnetic resonance imaging (MRI)-based diagnostics with therapeutic delivery and hyperthermia. However, their clinical translation requires sophisticated strategies to ensure precise delivery to the tumor site. This review examines innovative functionalization strategies to enhance the targeting and efficacy of SPMNPs. Specifically, it addresses the various strategies for coating magnetic nanoparticles with carbohydrates, including both covalent and non-covalent methods, and the subsequent functionalization of these glycoconjugates to exploit the unique biological environment of brain tumors. The use of glycoconjugates on the nanoparticle surface is a key strategy, leveraging the altered glycosylation patterns and overexpression of specific lectins on glioma cell surfaces to achieve highly selective cellular targeting. The review details the synergistic effect achieved by combining these functionalized nanoparticles with external magnetic field guidance. This combination provides a dual-action mechanism: the magnetic field actively guides the nanoparticles across the BBB and concentrates them within the tumor mass, while the carbohydrate coating ensures specific cellular uptake, thereby significantly improving local therapeutic concentration and minimizing systemic toxicity. The scope of this review includes the development and evaluation of carbohydrate-coated SPMNPs, outlining their optimized physicochemical properties for both in vitro and in vivo imaging and treatment of cancerous brain tissues. This comprehensive evaluation represents a critical advancement in biomedicine, aiming to improve the prognosis for patients with brain cancer through more precise and effective therapeutic interventions. Full article

Background/Objectives: The progression and dissemination of CRC are heavily influenced by a subpopulation of tumor cells known as CSCs. This study aimed to identify novel protein membrane antigens expressed by colorectal CSCs and the consequent development of targeted therapies based on monoclonal antibodies directed against the identified antigens. Methods: Integrated bioinformatics analyses were conducted using proprietary CSC gene expression profiles and public colon gene expression databases, leading to the identification of five plasma membrane proteins enriched in CSCs. Genetic immunization in rats was employed to generate monoclonal antibodies (mAbs) targeting these antigens. FGFR4 was prioritized due to its overexpression in colorectal tumors. Its function was characterized in vitro and in vivo through assays evaluating proliferation, colony formation, migration, and tumorigenicity. The anti-FGFR4 antibody 3B6 was selected based on its affinity and ability to inhibit FGFR4 signaling in CSCs. Its therapeutic potential was further assessed in xenograft models, and alterations in downstream signaling were analyzed via Western blot. Results: FGFR4 emerged as a key regulator of CRC CSC proliferation, migration, and tumorigenic capacity. The 3B6 antibody, a high-affinity FGFR4 binder, demonstrated robust in vitro inhibition of CSC features and significant antitumor effects in patient-derived xenograft models. Western blot analysis confirmed the modulation of FGFR4-driven signaling pathways, particularly those involved in epithelial–mesenchymal transition (EMT). Conclusions: This study successfully identified several CSC-selective membrane antigens that can become therapeutic targets in CRC. Among them, we focused on FGFR4 as a promising target and developed the anti-FGFR4 3B6 monoclonal antibody which offers potential for both diagnostic and therapeutic applications. Full article

Background: This study is part of the HEAL ITALIA partnership, funded by the National Recovery and Resilience Plan (PNRR) and the European Union. Heart failure (HF) is a serious health problem, with a reduced density of the β1-adrenergic receptor (β1-AR) in the myocardium as a hallmark. It is unclear whether this downregulation causes dysfunction or represents an epiphenomenon. Recent evidence implicates oxidative stress and mitochondrial signaling, particularly through the 18 kDa translocator protein (TSPO), in the regulation of the β1-AR, with possible modulation by estrogen. Objectives: To determine (1) the role of β1-AR desensitization in the onset and development of HF; (2) whether monocytes can represent a suitable ex vivo model for sex-oriented mechanistic studies in the cardiac field; (3) whether monocytes isolated from peripheral blood of patients can represent a diagnostic and/or therapy response biomarker by monitoring β1-AR density; (4) whether and how the mitochondrial receptor TSPO is involved in the β1-AR dysregulation observed in HF; and (5) whether the mechanisms linked to the onset of HF are regulated in a sex-specific manner through the effect of estrogen and/or the X chromosome on the expression of specific microRNAs. Methods: Using an integrated in vitro-ex vivo-in vivo methodological approach, we will evaluate the density of β1/β2-AR receptors, the downstream signaling (GRK2/β-arrestin), mitochondrial and redox parameters, and miRNA profiles in human monocytes and cardiomyocytes, and in mouse hearts after HF following pressure overload. Conclusions: The goal is to better understand the mechanisms underlying β1-AR desensitization, verify monocytes as peripheral markers of disease progression and response to therapy, and provide potentially useful information for the development of gender-specific therapies for heart failure. Full article

Liver fibrosis induced by Schistosoma japonicum Katsurada, 1904 (S. japonicum) infection lacks effective diagnostic markers and specific anti-fibrotic therapies. Although dysregulated iron homeostasis and ferroptosis pathways may contribute to its pathogenesis, the core regulatory mechanisms remain elusive. To unravel the ferroptosis-related molecular features, this study integrated transcriptomic datasets (GSE25713 and GSE59276) from S. japonicum-infected mouse livers. Following batch effect correction and normalization, ferroptosis-related differentially expressed genes (FRDEGs) were identified. Subsequently, core hub genes were screened through the construction of a protein–protein interaction (PPI) network, functional enrichment analysis, immune infiltration evaluation, and receiver operating characteristic (ROC) analysis. The expression patterns of these hub genes were further validated in an S. japonicum-infected mouse model using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The study identified 7 hub genes (Lcn2, Timp1, Cth, Cp, Hmox1, Cbs, and Gclc) as key regulatory molecules. Functional enrichment analysis revealed that these hub genes are closely associated with sulfur amino acid metabolism and oxidative stress responses. Specifically, key enzymes involved in cysteine and glutathione (GSH) synthesis (Cth, Cbs, Gclc) were consistently downregulated, suggesting a severe impairment of the host antioxidant defense capacity. Conversely, pro-fibrotic and pro-inflammatory markers (Timp1, Lcn2, Hmox1) were upregulated. This molecular pattern was significantly associated with a remodeled immune microenvironment, characterized by increased infiltration of neutrophils and eosinophils. In vivo validation confirmed the expression trends of 6 hub genes, corroborating the bioinformatics predictions, while the discrepancy in Cp expression highlighted the complexity of post-transcriptional regulation in vivo. The identified hub genes demonstrated excellent diagnostic potential, with Timp1 achieving an area under the curve (AUC) of 1.000. This study elucidates the molecular link between S. japonicum infection and the ferroptosis pathway, suggesting that these hub genes may drive liver fibrosis progression by regulating sulfur metabolism and the immune microenvironment. These findings offer potential diagnostic biomarkers and novel therapeutic targets for schistosomal liver fibrosis. Full article

Background: Cigarette burns represent a well-established forensic indicator of inflicted injury, frequently encountered in cases of domestic violence. Clinical significance: Their morphological consistency and anatomical distribution offer valuable elements for differentiating between intentional and accidental trauma. Case Presentation: In this study, we report the case of a 40-year-old woman who presented with multiple cutaneous lesions attributed to repeated assaults by her intimate partner. The forensic medical examination revealed five discrete scars characterized by sharply demarcated borders, circular to oval shapes, and dimensions ranging from 0.7 to 1.5 cm. These lesions were anatomically located in regions not typically accessible for self-infliction. To reinforce the diagnostic interpretation and assess reproducibility, a controlled experimental protocol was conducted using porcine skin matrices. Cigarette burns were recreated under variable conditions of contact pressure and exposure duration. The lesions produced on the biological substrate exhibited morphological features consistent with those observed in the patient, suggesting compatibility with cigarette-induced thermal injury. Conclusions: These findings provide circumstantial support for the forensic interpretation but must be considered within the limitations of the experimental model. This integrated approach underscores the relevance of combining clinical forensic documentation with experimental validation to support medico-legal conclusions in cases of suspected interpersonal violence. Full article

Next-generation therapeutic devices will rely on an intelligent integrated system that consolidates multiple functions into a single platform. These individual chemical components exhibit diverse physicochemical properties, demonstrating multifunctional characteristics. In this review, we focus on how the distinctive properties of upconversion nanoparticles (UCNPs), achieved via refined preparation methods, unlock novel functionalities in biomedical applications. Specifically, features such as near-infrared excitation, deep-tissue penetration, low autofluorescence, and tunable multicolor emission endow UCNPs with substantial potential in fields including deep-tissue imaging, targeted drug delivery, and photodynamic therapy. This article systematically reviews recent advances in the design and functionalization of UCNPs, elucidating their role in facilitating the development of integrated diagnostic and therapeutic platforms and fostering the establishment of intelligent responsive treatment systems. Finally, we address current technical challenges—including uniformity in large-scale production, long-term biosafety, and in vivo metabolic mechanisms—and provide insights into future interdisciplinary integration, clinical translation pathways, and their potential role in personalized medicine. Full article

We determined the in vivo absorption coefficient (μ_a) for 82 test subjects, all classified as Fitzpatrick skin phototypes II, III, IV, and V. Measurements were conducted using the integrating-sphere technique on the dorsal and palmar surfaces of the hand and the forearm. The reflectance data obtained were processed using the Inverse Adding Doubling algorithm to calculate the absorption coefficient. The mean values for this parameter ranged from 0.0132 mm⁻¹ to 0.1021 mm⁻¹ at a central wavelength of 624 nm. It was found that these parameters may be grouped into a distinct cohort, paving the way for studies and the design of light-based diagnostics and treatments better suited to the population in Mexico and Latin America. Full article

Background/Objectives: Despite clonal expansion during a primary immune response, or after subsequent antigen encounters, the frequency of memory B cells (B_mem) specific for an antigen remains low, making their detection difficult. However, unlike serum antibodies, which have a short half-life in vivo and thus require continuous replenishment to maintain stable titers, circulating B_mem are long-lived; they preserve immunological preparedness through their ability to rapidly engage in recall responses and differentiate into antibody-secreting cells (ASCs) upon antigen encounter. To this end, development of assays suited for the reliable detection of rare antigen-specific B_mem is critical and can provide insights into an individual’s antigen exposure history and immune status beyond that offered by traditional serum antibody measurements alone. Methods: ImmunoSpot^® has emerged as a suitable technique for the detection of individual antigen-specific B cells through visualizing their antibody-derived secretory footprints. Here, we report the theoretical and practical foundations for detecting rare antigen-specific B_mem in human peripheral blood mononuclear cells (PBMC). Leveraging the unique availability of verifiably naïve vs. antigen-experienced human samples, we used SARS-CoV-2 Spike (S-) and Nucleocapsid (NCAP) antigens to interrogate the presence of B_mem with these respective specificities. Results: While 100% diagnostic accuracy was achieved for both antigens, detection of NCAP-specific B_mem required reducing the lower detection limit of the standard assay. Specifically, this was achieved by testing a total of 2 million PBMC across multiple replicate assay wells and assessing the cumulative number of secretory footprints detected. Conclusion: The protocols described here should facilitate the reliable detection of ASCs present at varying precursor frequencies and serve as guidance for routine immune monitoring of rare B_mem with specificity for any antigen. Full article

Ionic liquids (ILs) have emerged as multifunctional compounds with low volatility, high thermal stability, and tunable solvation capabilities, making them highly promising for biomedical applications. First explored in the late 1990s and early 2000s for enhancing the thermal stability of enzymes, antimicrobial agents, and controlled release systems, ILs have since gained significant attention in drug delivery, antimicrobial treatments, medical imaging, and biosensing. This review examines the diverse functions of ILs in contemporary therapeutics and diagnostics, highlighting their transformative capabilities in improving drug solubility, bioavailability, transdermal permeability, and pathogen inactivation. In drug delivery, ILs improve solubility of bioactive compounds, with several IL formulations achieving substantial solubility enhancements for poorly soluble drugs. Bio-ILs, in particular, show promise in enhancing drug delivery systems, such as improving transdermal permeability. ILs also exhibit significant antimicrobial and antiviral activity, offering new avenues for combating resistant pathogens. Despite their broad potential, challenges such as cytotoxicity, long-term metabolic effects, and the stability of ILs in physiological conditions persist. While much research has focused on their physicochemical properties, biological activity and in vivo studies are still underexplored. The future directions for ILs in biomedical applications include the development of bioengineered ILs and hybrid ILs, combining functional components like nanoparticles and polymers to create multifunctional materials. These ILs, derived from renewable resources, show great promise in personalized medicine and clinical applications. Further research is necessary to evaluate their pharmacokinetics, biodistribution, and long-term safety to fully realize their biomedical potential. This study emphasizes the potential of ILs to transform therapeutic and diagnostic technologies by highlighting present shortcomings and offering pathways for clinical translation, while also debating the need for continuous research to fully utilize their biomedical capabilities. Full article

Subcutaneous mycoses are a heterogeneous group of chronic fungal infections, usually acquired through traumatic inoculation of environmental fungi and particularly severe in immunocompromised and critically ill patients. These infections involve pathogens with marked morphological and physiopathological diversity, resulting in significant diagnostic and therapeutic challenges. Conventional treatment relies on systemic antifungals such as amphotericin B, itraconazole, and other azoles; however, these therapies are often limited by poor tissue penetration, adverse effects, and prolonged treatment regimens, especially in vulnerable patient populations. In this context, nanodrugs have emerged as promising alternatives by improving solubility, stability, bioavailability, and targeted delivery to infection sites. This review conducted a comprehensive literature search in PubMed, SciELO, ScienceDirect, Web of Science, and Scopus, identifying 31 eligible studies that developed and evaluated antifungal nanosystems using in vitro, ex vivo, and/or in vivo models. Quantitative outcomes included minimum inhibitory concentration (MIC), colony-forming units (CFU), inhibition halo diameter, and survival assays. Overall, the evidence indicates that several nanosystems may overcome key pharmacological limitations of conventional antifungals and enhance therapeutic outcomes. Nevertheless, important translational challenges remain, including toxicity, long-term safety, scalability, and regulatory approval, which must be addressed before clinical implementation. Full article