Search Results (27)

Dynamic monitoring of hemostatic equilibrium is indispensable for clinical safety in high-risk scenarios, while current clinical methods are limited by sample volume, detection speed, and physiological relevance. These shortcomings underscore the demand for novel sensing platforms. Optical biosensors, leveraging label-free detection, rapid response, and multi-level characterization, could serve as a transformative solution for decentralized and point-of-care monitoring. This review systematically summarizes advances in optical coagulation testing, encompassing light transmission aggregometry, laser speckle rheology, optical coherence tomography/elastography, optic–acoustic coupled methods, and fluorescence biosensing. These technologies complementarily capture structural and mechanical and some molecular and cellular dynamics of coagulation, bridging gaps in traditional assays. Despite promising preclinical and clinical correlations, translation barriers persist in lack of standardization of metrics, interference mitigation, and multi-center validation in diverse patient cohorts. Future development of optical biosensing platforms for coagulation testing should focus on modular integration, AI-aided interference correction, and microfluidic miniaturization to realize actionable, real-time coagulation assessment. Optical biosensors hold unparalleled potential to transform hemostatic monitoring from static endpoint testing to dynamic, interpretable evaluation, guiding personalized clinical decisions. Full article

Background: Plasma fibrinogen (FIB) levels exhibit a significant elevation during the acute phase of ischemic stroke (IS), and their dynamic fluctuations serve as important biomarkers for stroke onset, disease progression, and long-term prognosis. Tong-Qiao-Huo-Xue Decoction (TQHXD) is highly effective in treating blood stasis syndromes affecting the head and face. Nevertheless, the association between TQHXD and FIB in the underlying mechanism of treating IS warrants further investigation. Methods: Proteomics analysis predicted the potential therapeutic targets of TQHXD for IS. An in vivo model of middle cerebral artery occlusion followed by reperfusion (MCAO/R) was created in mice. To explore the interaction between FIB and NLRP3, as well as to verify the particular healing outcomes of TQHXD. Results: An increased blood–brain barrier (BBB) permeability was observed after MCAO/R, accompanied by substantial accumulation of FIB in the brain. In vivo experiments demonstrated that FIB triggered the activation of the NLRP3 inflammasome in microglia. Proteomic analysis revealed a significant increase in FIB levels following model induction, which were markedly reduced after treatment with TQHXD; KEGG pathway enrichment analysis indicated that these changes were primarily associated with the NOD-like receptor signaling pathway. Laser speckle contrast imaging showed that TQHXD treatment significantly improved cerebral blood flow and attenuated brain injury in mice. Fluorescence imaging, ELISA, and Western blotting results collectively demonstrated that TQHXD effectively reduced FIB accumulation and suppressed NLRP3 inflammasome activation. MD and pull-down experiments further demonstrated a strong interaction strength between FIB and NLRP3. Conclusions: FIB accumulates in the ischemic penumbra following CIRI, while TQHXD can effectively down-regulate FIB expression and inhibit NLRP3 inflammasome activation to mitigate CIRI. These findings provide a novel theoretical foundation and treatment direction for stroke management in clinical settings. Full article

Background: While intravenous administration of nanoparticles (NPs) is effective for targeting the lungs and liver, directing them to other organs and tissues remains challenging. Methods: Here, we report alternative administration routes that improve organ-specific accumulation of poly (lactic-co-glycolic acid) (PLGA) NPs (100 nm, negatively charged) loaded with the near-infrared dye Cyanine 7 (Cy7). NP cytotoxicity was evaluated in HEK293, mMSCs, C2C12, L929, and RAW264.7 cells. Hemocompatibility was assessed using WBCs and RBCs. NPs were administered via the tail vein, carotid, renal, and femoral arteries in BALB/c mice. Administration safety was evaluated by laser speckle contrast imaging and histological analysis. NP biodistribution and accumulation were assessed using in vivo and ex vivo fluorescence tomography and confocal microscopy of cryosections. Results: PLGA-Cy7 NPs demonstrate low cytotoxicity even at high doses and exhibit good hemocompatibility. Administration of NPs through the mouse carotid, renal, and femoral arteries significantly increases accumulation in the target ipsilateral brain hemisphere (31.7-fold) and salivary glands (28.3-fold), kidney (13.7-fold), and hind paw (3.6-fold), respectively, compared to intravenous administration. Injection of NPs through arteries supplying the target organs and tissues does not result in significant changes in blood flow, morphological alterations, or irreversible embolization of vessels, provided the procedure is performed correctly and the optimal dosage is used. Conclusions: These results highlight the potential of intra-arterial delivery of NPs for organ-specific drug targeting, underscoring the synergistic impact of advances in materials science, minimally invasive endovascular surgery, and nanomedicine. Full article

This study presents the results of over thirty years of non-destructive testing (NDT) in a historic church, providing an unprecedented time analysis of the structural and material integrity of the building and its works of art. During this time, the church has undergone several restorations and two major seismic events. The diagnostics, which include a calibrated mix of established and advanced micro-destructive and non-destructive (NDT) techniques such as X-ray fluorescence, holographic interferometry, electronic speckle pattern interferometry (ESPI), infrared thermography, and IR reflectography, provide critical insights into the impact of the restoration interventions and the earthquakes on the church’s artistic heritage. The results indicate varying degrees of effectiveness of the restoration efforts, highlighting both areas of successful conservation and emerging vulnerabilities. This long-term study highlights the importance of continuous monitoring and its integration with NDT in identifying the effects of time and strong events occurring during the life of artworks that influence their state of conservation. Full article

Objectives: This review aims to explore recent advancements in optical imaging techniques for monitoring the viability of Deep Inferior Epigastric Perforator (DIEP) flap reconstruction. The objectives include highlighting the principles, applications, and clinical utility of optical imaging modalities such as near-infrared spectroscopy (NIRS), indocyanine green (ICG) fluorescence angiography, laser speckle contrast imaging (LSCI), hyperspectral imaging (HSI), dynamic infrared thermography (DIRT), and short-wave infrared thermography (SWIR) in assessing tissue perfusion and oxygenation. Additionally, this review aims to discuss the potential of these techniques in enhancing surgical outcomes by enabling timely intervention in cases of compromised flap perfusion. Materials and Methods: A comprehensive literature review was conducted to identify studies focusing on optical imaging techniques for monitoring DIEP flap viability. We searched PubMed, MEDLINE, and relevant databases, including Google Scholar, Web of Science, Scopus, PsycINFO, IEEE Xplore, and ProQuest Dissertations & Theses, among others, using specific keywords related to optical imaging, DIEP flap reconstruction, tissue perfusion, and surgical outcomes. This extensive search ensured we gathered comprehensive data for our analysis. Articles discussing the principles, applications, and clinical use of NIRS, ICG fluorescence angiography, LSCI, HSI, DIRT, and SWIR in DIEP flap monitoring were selected for inclusion. Data regarding the techniques’ effectiveness, advantages, limitations, and potential impact on surgical decision-making were extracted and synthesized. Results: Optical imaging modalities, including NIRS, ICG fluorescence angiography, LSCI, HSI, DIRT, and SWIR offer a non- or minimal-invasive, real-time assessment of tissue perfusion and oxygenation in DIEP flap reconstruction. These techniques provide objective and quantitative data, enabling surgeons to monitor flap viability accurately. Studies have demonstrated the effectiveness of optical imaging in detecting compromised perfusion and facilitating timely intervention, thereby reducing the risk of flap complications such as partial or total loss. Furthermore, optical imaging modalities have shown promise in improving surgical outcomes by guiding intraoperative decision-making and optimizing patient care. Conclusions: Recent advancements in optical imaging techniques present valuable tools for monitoring the viability of DIEP flap reconstruction. NIRS, ICG fluorescence angiography, LSCI, HSI, DIRT, and SWIR offer a non- or minimal-invasive, real-time assessment of tissue perfusion and oxygenation, enabling accurate evaluation of flap viability. These modalities have the potential to enhance surgical outcomes by facilitating timely intervention in cases of compromised perfusion, thereby reducing the risk of flap complications. Incorporating optical imaging into clinical practice can provide surgeons with objective and quantitative data, assisting in informed decision-making for optimal patient care in DIEP flap reconstruction surgeries. Full article

Nuclear speckles are compartments enriched in splicing factors present in the nucleoplasm of eucaryote cells. Speckles have been studied in mammalian culture and tissue cells, as well as in some non-mammalian vertebrate cells and invertebrate oocytes. In mammals, their morphology is linked to the transcriptional and splicing activities of the cell through a recruitment mechanism. In rats, speckle morphology depends on the hormonal cycle. In the present work, we explore whether a similar situation is also present in non-mammalian cells during the reproductive cycle. We studied the speckled pattern in several tissues of a viviparous reptile, the lizard Sceloporus torquatus, during two different stages of reproduction. We used immunofluorescence staining against splicing factors in hepatocytes and oviduct epithelium cells and fluorescence and confocal microscopy, as well as ultrastructural immunolocalization and EDTA contrast in Transmission Electron Microscopy. The distribution of splicing factors in the nucleoplasm of oviductal cells and hepatocytes coincides with the nuclear-speckled pattern described in mammals. Ultrastructurally, those cell types display Interchromatin Granule Clusters and Perichromatin Fibers. In addition, the morphology of speckles varies in oviduct cells at the two stages of the reproductive cycle analyzed, paralleling the phenomenon observed in the rat. The results show that the morphology of speckles in reptile cells depends upon the reproductive stage as it occurs in mammals. Full article

SARS-CoV-2 infection remains a global burden. Despite intensive research, the mechanism and dynamics of early viral replication are not completely understood, such as the kinetics of the formation of genomic RNA (gRNA), sub-genomic RNA (sgRNA), and replication centers/organelles (ROs). We employed single-molecule RNA-fluorescence in situ hybridization (smRNA-FISH) to simultaneously detect viral gRNA and sgRNA and immunofluorescence to detect nsp3 protein, a marker for the formation of RO, and carried out a time-course analysis. We found that single molecules of gRNA are visible within the cytoplasm at 30 min post infection (p.i.). Starting from 2 h p.i., most of the viral RNA existed in clusters/speckles, some of which were surrounded by single molecules of sgRNA. These speckles associated with nsp3 protein starting at 3 h p.i., indicating that these were precursors to ROs. Furthermore, RNA replication was asynchronous, as cells with RNA at all stages of replication were found at any given time point. Our probes detected the SARS-CoV-2 variants of concern, and also suggested that the BA.1 strain exhibited a slower rate of replication kinetics than the WA1 strain. Our results provide insights into the kinetics of SARS-CoV-2 early post-entry events, which will facilitate identification of new therapeutic targets for early-stage replication to combat COVID-19. Full article

Atmospheric CO₂ levels have been increasing, and these changes may result in differential adaptive responses in both genera and species and highlight the need to increase carbon sequestration. Ecophysiological and morphological responses of four early-successional deciduous species were examined under ambient CO₂ (aCO₂, 400 ppm) and elevated CO₂ (eCO₂, 800 ppm) treatments. The four species, all of which are used in restoration, were Alnus viridis subsp. crispa (Ait.) Turrill (green alder), A. incana subsp. rugosa (Du Roi) R.T. Clausen (speckled alder), Betula populifolia (Marshall) (gray birch), and B. papyrifera (Marshall) (white birch); all are from the same phylogenetic family, Betulaceae. We examined biochemical efficiencies, gas exchange, chlorophyll fluorescence, chlorophyll concentrations, foliar nitrogen (N), and growth traits. A general linear model, analysis of variance, was used to analyze the functional carbon efficiency and growth differences, if any, among genera, species, and provenances (only for growth traits). The alders had greater biochemical efficiency traits than birches, and alders upregulated these traits, whereas birches mostly downregulated these traits in response to eCO₂. In response to eCO₂, assimilation either remained the same or was upregulated for alders but downregulated for birches. Stomatal conductance was downregulated for all four species in response to eCO₂. Intrinsic water use efficiency was greater for alders than for birches. Alders exhibited a consistent upregulation of stem dry mass and height growth, whereas birches were somewhat lower in height and stem dry mass in response to eCO₂. Foliar N played an important role in relation to ecophysiological traits and had significant effects relative to genus (alders > birches) and CO₂ (aCO₂ > eCO₂), and a significant genus × CO₂ interaction, with alders downregulating foliar N less than did birches. Covariate analysis examining carbon efficiency traits in relation to foliar N showed clear functional responses. Both species in both genera were consistent in their ecophysiological and morphological responses to CO₂ treatments. There was supporting evidence that assimilation was sink-driven, which is related to a plant organ’s ability to continue to grow and incorporate assimilates. The alders used in this study are actinorhizal, and the additional available foliar N, paired with increased stem dry mass sink activity, appeared to be driving upregulation of the carbon efficiencies and growth in response to eCO₂. Alders’ greater carbon efficiencies and carbon sequestration in impoverished soils demonstrate that alders, as opposed to birches, should be used to accelerate ecological restoration in a world of increasing atmospheric CO₂. Full article

The diagnosis of systemic autoimmune rheumatic disease (SARD) or its exclusion is carried out taking into account the results of immunological studies, primarily antinuclear antibodies (ANA) and specific autoantibodies. Often, during ANA analysis via indirect immunofluorescence reaction on cellular and tissue substrates, a dense fine speckled 70 (DFS70) fluorescence pattern is observed. Studies on the diagnostic significance of antibodies to anti-DFS70 allow for optimizing the stepwise diagnosis of SARD. Currently, a two-step strategy for laboratory diagnostic investigation is recommended: in the first step, ANA screening is performed, and in the second step, patients with positive results undergo confirmatory tests to detect specific antibodies against individual nuclear antigens. The detection of anti-DFS70 in ANA-seropositive patients without clinical and/or other specific serological markers characteristic of a particular disease within the SARD group may be considered a negative prognostic marker. Also, in the process of decision making in clinical practice, we should remember that anti-DFS70 can be found in the blood of patients with a different, non-SARD pathology and that most people showing anti-DFS70 are healthy individuals. Full article

We propose a novel hybrid FPP-DIC technique to measure an object’s shape and deformation in 3D simultaneously by using a single 3CCD color camera, which captures the blue fringe patterns and red fluorescent speckles within the same image. Firstly, red fluorescent speckles were painted on the surface of the specimen. Subsequently, 12 computer-generated blue fringe patterns with a black background were projected onto the surface of the specimen using a DLP projector. Finally, both the reference and deformed images with three different frequencies and four shifted phases were captured using a 3CCD camera. This technique employed a three-chip configuration in which red–green–blue chips were discretely integrated in the 3CCD color camera sensor, rendering independent capture of RGB information possible. Measurement of out-of-plane displacement was carried out through the implementation of Fringe Projection Profilometry (FPP), whereas the in-plane displacement was evaluated using a 2D Digital Image Correlation (DIC) method by leveraging a telecentric-lens-based optical system. In comparison to the traditional FPP-DIC hybrid methodology, the present approach showed a lower incidence of crosstalk between the fringe patterns and speckle patterns while also offering a corrective for the coupling of the in-plane displacement and out-of-plane displacement. Experimental results for the in-plane cantilever beam and out-of-plane disk comparisons with the traditional 3D-DIC method indicated that the maximum discrepancy obtained between FPP-DIC and 3D-DIC was 0.7 μm and 0.034 mm with different magnifications, respectively, validating the effectiveness and precision of the novel proposed FPP-DIC method. Full article

Signaling and detoxification of Reactive Oxygen Species (ROS) are important patho-physiologcal processes. Despite this, we lack comprehensive information on individual cells and cellular structures and functions affected by ROS, which is essential to build quantitative models of the effects of ROS. The thiol groups from cysteines (Cys) in proteins play a major role in redox defense, signaling, and protein function. In this study, we show that the proteins in each subcellular compartment contain a characteristic Cys amount. Using a fluorescent assay for -SH in thiolate form and amino groups in proteins, we show that the thiolate content correlates with ROS sensitivity and signaling properties of each compartment. The highest absolute thiolate concentration was found in the nucleolus, followed by the nucleoplasm and cytoplasm whereas protein thiolate groups per protein showed an inverse pattern. In the nucleoplasm, protein reactive thiols concentrated in SC35 speckles, SMN, and the IBODY that accumulated oxidized RNA. Our findings have important functional consequences, and explain differential sensitivity to ROS. Full article

The objective of this study is to use imaging and histopathological analysis to characterize and monitor microvascular responses to photodynamic therapy (PDT). In vivo chicken chorioallantoic membranes (CAMs) and a stimulated malignant oral lesions animal model were used to determine the blood flow and the biological activities of Photofrin^® (2.5 mg/kg) exposed to different laser power densities at 630 nm. The vascular changes, the velocity of the blood flow, the speckle flow index (SFI) of fluorescence changes, and ultrastructure damage in the microvasculature before and after PDT were recorded. The subcellular localization of Photofrin^® revealed satisfactory uptake throughout the cytoplasm of human red blood cells at 10 s and 20 s before PDT. The mean blood-flow velocities of the veins and arteries were 500 ± 40 and 1500 ± 100 μm/s, respectively. A significant decrease in the velocities of the blood flow in the veins and arteries was detected in the CAM model after PDT. The veins and arteries of CAMs, exposed to the power densities of 80, 100, and 120 mW/cm², had average blood-flow velocities of 100 ± 20, 60 ± 10, and 0 μm/s and 300 ± 50, 150 ± 30, and 0 μm/s, respectively. In the stimulated malignant oral lesions animal model, the treated tumors exhibited hemorrhage and red blood cell extravasation after PDT. The oxyhemoglobin and total hemoglobin levels decreased, which resulted in a decrease in tissue oxygen saturation, while the deoxyhemoglobin levels increased. PDT using Photofrin^® has the ability to cause the destruction of the targeted microvasculature under nonthermal mechanisms selectively. Full article

Background: The presence of Antinuclear antibodies/Dense Fine Speckled 70 (ANA/DFS70) has been proposed as a negative biomarker in the process of exclusion of systemic autoimmune/autoinflammatory rheumatic diseases (SARD). The purpose was to evaluate and characterize ANA/DFS70 patients in a large Colombian population with SARD; rheumatoid arthritis (RA), Psoriasis (PsO), Undifferentiated connective tissue disease (UCTD), first-degree relatives of (FDR), and healthy controls (HC). Methods: ANA determination was performed using indirect immunofluorescence. Samples with positive dense fine granular staining in the nucleoplasm of the interphase cell (AC2) fluorescence were confirmed with CytoBead/ANA and ANA/modified (Knocked out for the PSPI1 gen). Results: 530 mestizo Colombian participants were included. ANA/DFS70 antibody positivity in the whole group was 2.3%, and 0.8% in SARD; no RA patients were positive. ANA/DFS70 positives in UCTD were three women; the average time of evolution of the disease was 9.4 years. The most frequent clinical findings were arthralgias, non-erosive arthritis, and Raynaud’s phenomenon. The PsO positive was a woman with C-reactive protein (CRP) positivity and a negative erythrocyte sedimentation rate (ESR) without any other positive autoantibody or extracutaneous manifestation. FDR and HC positives were 7/8 women. All were negative for other autoantibodies. Conclusions: ANA/DFS70 autoantibodies were present in Colombian patients with SARD at a shallow frequency, they were more prevalent in healthy individuals. Full article

A fundamental limitation of the spectral response of laser-pumped fluorescent nanostructured media was considered in terms of a probabilistic model establishing the relationship between the enhancement factor of a spectral quality and characteristic propagation and amplification scales of pump light and fluorescence in the medium. It was shown that the minimum spectral width of fluorescent response of the pumped medium is limited by competition between the stimulated emission and radiation losses in microscopic fluorescence emitters associated with the speckles randomly modulating the pumping light field. Theoretical results were compared to the experimental data on the spectral properties of the fluorescent response of laser-pumped nanostructured “anatase nanoparticles—dye solutions” systems with various structural and optical properties. Rhodamine 6G and 4-(dicyanomethylene)-2-methyl-6-(p-dimethylaminostyryl)-4H-pyran (DCM) were applied as fluorescent components in the examined systems with various scatter volume fractions, which were pumped by pulse-periodic laser radiation with various intensities at 532 nm. Comparison showed a fair agreement between the theoretical and experimental results. Full article

Nuclear speckles are nuclear bodies containing RNA-binding proteins as well as RNAs including long non-coding RNAs (lncRNAs). Maternally expressed gene 3 (MEG3) is a nuclear retained lncRNA found to associate with nuclear speckles. To understand the association dynamics of MEG3 lncRNA with nuclear speckles in living cells, we generated a fluorescently tagged MEG3 transcript that could be detected in real time. Under regular conditions, transient association of MEG3 with nuclear speckles was observed, including a nucleoplasmic fraction. Transcription or splicing inactivation conditions, known to affect nuclear speckle structure, showed prominent and increased association of MEG3 lncRNA with the nuclear speckles, specifically forming a ring-like structure around the nuclear speckles. This contrasted with metastasis-associated lung adenocarcinoma (MALAT1) lncRNA that is normally highly associated with nuclear speckles, which was released and dispersed in the nucleoplasm. Under normal conditions, MEG3 dynamically associated with the periphery of the nuclear speckles, but under transcription or splicing inhibition, MEG3 could also enter the center of the nuclear speckle. Altogether, using live-cell imaging approaches, we find that MEG3 lncRNA is a transient resident of nuclear speckles and that its association with this nuclear body is modulated by the levels of transcription and splicing activities in the cell. Full article