Flow Imaging Research Articles

Understanding the relationship between silicone implants, tumor antigens, and breast cancer risk: An immunological study in rats.

This study aimed to investigate the effects of silicone implants on the incidence of breast cancer in rats, as well as their impact on immune surveillance mechanisms. Female SD rats were divided into three groups: a Placebo Surgery Group (PSG), a Thoracic Implant Group (TIG), and a Back Implant Group (BIG). Following the corresponding surgical procedures, we measured Secretoglobin Family 2A, Member 2(SCGB2A2) and Mucin-1 (MUC1) antigen levels using ELISA, and statistical analyses were conducted to evaluate immune responses. The N-Methyl-N-Nitrosourea(MNU)-induced breast cancer model and pathological analyses indicated that the incidence of breast cancer in the thoracic implant group was lower, suggesting that silicone implants may reduce the risk of breast cancer. Additionally, laser speckle blood flow imaging and immunohistochemical analysis revealed blood perfusion in the implant capsule area and an active response of immune cells, indicating that immune surveillance may exert local effects. These findings provide the first evidence of a relationship between tumor antigens, silicone implants, and breast cancer incidence, offering a new immunological perspective on the safety of silicone implants.

Effect of electroacupuncture on vascular remodeling in rats with cerebral ischemia by regulating irisin based on VEGF/Akt/eNOS signaling pathway.

This study aimed to explore the cumulative effects and expression patterns of electroacupuncture (EA) on irisin secretion, observe the effects of EA on the recovery of neurobehavioral function and vascular remodeling after cerebral ischemia, and elucidate the mechanism by which EA promotes vascular remodeling by regulating irisin expression. A rat model of left middle cerebral artery occlusion (MCAO) was prepared, and EA was performed. Tissue distribution and expression of irisin were determined by immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and Western blotting. Type III fibronectin domain protein 5-silenced adeno-associated virus (rAAV-shFNDC5) was injected into the lateral ventricle as a control. Neurobehavioral function was evaluated using 2,3,5-triphenyltetrazolium chloride (TTC) staining and behavioral experiments, while vascular remodeling was evaluated using laser speckle blood flow imaging, and the expressions of irisin and vascular remodeling-related factors were measured by ELISA and Western blotting. The number of FNDC5-positive neurons, fluorescence intensity, and irisin expression reached their maximum increase after 7 days of EA treatment. In addition, the EA group exhibited a significant reduction in cerebral infarct volume and impairment of neurobehavioral function, an increase in cerebral blood flow and microvascular diameter on the ischemic side, and significantly higher expression levels of FNDC5, brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), protein kinase B (Akt), and endothelial nitric oxide synthase (eNOS). However, rAAV-shFNDC5 significantly weakened the therapeutic effects of EA. EA upregulated irisin expression, reaching a peak after 7 days of EA and then stabilizing. EA facilitated vascular remodeling after cerebral ischemia, and this might be associated with the activation of the irisin-mediated VEGF/Akt/eNOS signaling pathway.

Imaging flow cytometry reveals LPS-induced changes to intracellular intensity and distribution of α-synuclein in a TLR4-dependent manner in STC-1 cells

BackgroundParkinson’s disease is a chronic neurodegenerative disorder, where pathological protein aggregates largely composed of phosphorylated α-synuclein are implicated in disease pathogenesis and progression. Emerging evidence suggests that the interaction between pro-inflammatory microbial factors and the gut epithelium contributes to α-synuclein aggregation in the enteric nervous system. However, the cellular sources and mechanisms for α-synuclein pathology in the gut are still unclear. MethodsThe STC-1 cell line, which models an enteroendocrine population capable of communicating with the microbiota, immune and nervous systems, was treated with a TLR4 inhibitor (TAK-242) prior to microbial lipopolysaccharide (LPS) exposure to investigate the role of TLR4 signalling in α-synuclein alterations. Antibodies targeting the full-length protein (α-synuclein) and the Serine-129 phosphorylated form (pS129) were used. Complex, multi-parametric image analysis was conducted through confocal microscopy (with Zen 3.8 analysis) and imaging flow cytometry (with IDEAS® analysis). ResultsConfocal microscopy revealed heterogenous distribution of α-synuclein and pS129 in STC-1 cells, with prominent pS129 staining along cytoplasmic processes. Imaging flow cytometry further quantified the relationship between various α-synuclein morphometric features. Thereafter, imaging flow cytometry demonstrated a dose-specific effect of LPS, where the low (8 μg/mL), but not high dose (32 μg/mL), significantly altered measures related to α-synuclein intensity, distribution, and localisation. Pre-treatment with a TLR4 inhibitor TAK-242 alleviated some of these significant alterations. ConclusionThis study demonstrates that LPS-TLR4 signalling alters the intracellular localisation of α-synuclein in enteroendocrine cells in vitro and showcases the utility of combining imaging flow cytometry to investigate subtle protein changes that may not be apparent through confocal microscopy alone. Further investigation is required to understand the apparent dose-dependent effects of LPS on α-synuclein in the gut epithelium in healthy states as well as conditions such as Parkinson’s disease.

Programmable scanning diffuse speckle contrast imaging of cerebral blood flow.

Cerebral blood flow (CBF) imaging is crucial for diagnosing cerebrovascular diseases. However, existing large neuroimaging techniques with high cost, low sampling rate, and poor mobility make them unsuitable for continuous and longitudinal CBF monitoring at the bedside. We aimed to develop a low-cost, portable, programmable scanning diffuse speckle contrast imaging (PS-DSCI) technology for fast, high-density, and depth-sensitive imaging of CBF in rodents. The PS-DSCI employed a programmable digital micromirror device (DMD) for remote line-shaped laser (785nm) scanning on tissue surface and synchronized a 2D camera for capturing boundary diffuse laser speckle contrasts. New algorithms were developed to address deformations of line-shaped scanning, thus minimizing CBF reconstruction artifacts. The PS-DSCI was examined in head-simulating phantoms and adult mice. The PS-DSCI enables resolving intralipid particle flow contrasts at different tissue depths. In vivo experiments in adult mice demonstrated the capability of PS-DSCI to image global/regional CBF variations induced by 8% inhalation and transient carotid artery ligations. Compared with conventional point scanning, line scanning in PS-DSCI significantly increases spatiotemporal resolution. The high sampling rate of PS-DSCI is crucial for capturing rapid CBF changes while high spatial resolution is important for visualizing brain vasculature.

Ide Copy Number Variant Does Not Influence Stroke Severity in 2 C57BL/6J Mouse Models nor in Humans: An Exploratory Study.

Contrary to the common belief, the most commonly used laboratory C57BL/6J mouse inbred strain presents a distinctive genetic and phenotypic variability, and for several traits, the genotype-phenotype link remains still unknown. Recently, we characterized the most important stroke survival factor such as brain collateral plasticity in 2 brain ischemia C57BL/6J mouse models (bilateral common carotid artery stenosis and middle cerebral artery occlusion) and observed a Mendelian-like fashion of inheritance of the posterior communicating artery (PcomA) patency. Interestingly, a copy number variant (CNV) spanning Ide locus was reported to segregate in an analogous Mendelian-like pattern in the C57BL/6J colonies of the Jackson Laboratory. Given IDE critical role in vascular plasticity, we hypothesized Ide CNV may have explained PcomA variability in C57BL/6J inbred mice. We applied a combination of techniques (T2-weighted magnetic resonance imaging, time-of-flight angiography, cerebral blood flow imaging, and histology) to characterize the collaterome in 77 C57BL/6J bilateral common carotid artery stenosis, middle cerebral artery occlusion, naive, and sham mice and performed on these Taqman genotyping, exome sequencing, and RNA sequencing. We then investigated the hypothesis that IDE structural variants (CNVs, gain/loss of function mutations) may have influenced the cerebrovascular phenotype in a large cohort of 454 040 cases and controls (UK Biobank, Genomics England). We detected an Ide CNV in a bilateral common carotid artery stenosis mouse with 2 patent PcomAs (minor allele frequency, 1.3%), not segregating with the PcomA patency phenotype. In addition, 2 heterozygous IDE CNVs, resulting in loss of function were found in 1 patient with hereditary ataxia, a patient with hereditary congenital heart disease, and 2 healthy individuals (minor allele frequency 9×10-6). Moreover, we report 4 IDE loss of function point mutations (p.Leu5X, p.Met394ValfsX29, p.Pro14SerfsX26, p.Leu889X, minor allele frequency 0.02%) present also in controls or inherited from healthy parents. Ide CNV and loss of function variants are rare, do not crucially influence PcomA variability in C57BL/6J inbred mice, and do not cause a vascular phenotype in humans.

Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization

The process of viral entry into host cells is crucial for the establishment of infection and the determination of viral pathogenicity. A comprehensive understanding of entry pathways is fundamental for the development of novel therapeutic strategies. Standard techniques for investigating viral entry include confocal microscopy and flow cytometry, both of which provide complementary qualitative and quantitative data. Imaging flow cytometry, which integrates the advantages of both methodologies, offers significant potential in virological studies. In this investigation, we employed imaging flow cytometry coupled with immunostaining to monitor the entry of equine arteritis virus EAV into Vero cells via the endosomal trafficking route. Analysis provided an insight into the early infection dynamics across thousands of cells, revealing statistically significant alterations in internalization and uncoating process. Moreover, we evaluated the effectiveness of two inhibitors targeting cellular factors involved in facilitating viral entry: ammonium chloride, which disrupts endocytosis, and camostat mesylate, which inhibits the activity of serine proteases. The results demonstrated a clear distinction between effective and ineffective inhibitors. This study highlighted the potential of imaging flow cytometry to advance the study of viral entry and the evaluation of antiviral agents.

Extracellular vesicles in ageing cold-stored whole blood may not compensate for the decreasing haemostatic function in vitro.

Extracellular vesicles (EVs) have procoagulative properties. As EVs are known to accumulate in stored blood products, we compared the EV content and coagulation capacity of leukoreduced cold-stored whole blood (CSWB) with current prehospital and in-hospital component therapies to understand the role of EVs in the haemostatic capacity of ageing CSWB. Blood was obtained from 12 O RhD-positive male donors. CSWB was compared with in-hospital component therapy of red blood cells (RBCs), OctaplasLG and buffy-coat platelets and prehospital component therapy of RBC and lyophilized plasma. Samples were drawn on Days 1 and 14 of CSWB and RBC cold storage. Blood count, haemolysis markers, rotational thromboelastometry, sonorheometry and thrombin generation were analysed. EVs were analysed using nanoparticle tracking analysis and cellular origin was determined using imaging flow cytometry. There was a trend towards increased production of both platelet and RBC-derived EVs during CSWB storage. Particle count increased during storage, whereas thrombin generation slowed down and in viscoelastic assays, clotting times prolonged, clot formation became impaired, and stiffness of the resulting clot decreased. Both platelet and RBC-derived EVs increased in number in CSWB during storage. This did not appear to compensate for the in vitro decreasing haemostatic capacity of ageing CSWB, suggesting EVs produced during storage may not have active procoagulative effects, but rather reflect the ageing of blood cells.

Protocol for assessing and visualizing cell microaggregate formation in whole blood by imaging flow cytometry.

Blood cell aggregates are clinically useful biomarkers in a number of medical disorders. This protocol provides accurate and quantitative analysis of cell aggregates using a small volume of whole blood and imaging flow cytometry. We describe steps for sample collection, staining, and measurement. We then detail gating procedures and analysis of cell morphology. Sample preparation artifacts, activation, and morphological changes of cells are mitigated by omitting erythrocyte lysis and leukocyte isolation while maintaining high-throughput accurate imaging of leukocytes and platelets.

A deep learning approach for early prediction of breast cancer neoadjuvant chemotherapy response on multistage bimodal ultrasound images

Neoadjuvant chemotherapy (NAC) is a systemic and systematic chemotherapy regimen for breast cancer patients before surgery. However, NAC is not effective for everyone, and the process is excruciating. Therefore, accurate early prediction of the efficacy of NAC is essential for the clinical diagnosis and treatment of patients. In this study, a novel convolutional neural network model with bimodal layer-wise feature fusion module (BLFFM) and temporal hybrid attention module (THAM) is proposed, which uses multistage bimodal ultrasound images as input for early prediction of the efficacy of neoadjuvant chemotherapy in locally advanced breast cancer (LABC) patients. The BLFFM can effectively mine the highly complex correlation and complementary feature information between gray-scale ultrasound (GUS) and color Doppler blood flow imaging (CDFI). The THAM is able to focus on key features of lesion progression before and after one cycle of NAC. The GUS and CDFI videos of 101 patients collected from cooperative medical institutions were preprocessed to obtain 3000 sets of multistage bimodal ultrasound image combinations for experiments. The experimental results show that the proposed model is effective and outperforms the compared models. The code will be published on the https://github.com/jinzhuwei/BLTA-CNN.

Microvascular blood flow ultrasound imaging with microbubble-based H-Scan technology.

Ultrasound blood flow imaging plays a crucial role in the diagnosis of cardiovascular and cerebrovascular diseases. Conventional ultrafast ultrasound plane-wave imaging techniques have limited capabilities in microvascular imaging. To enhance the quality of blood flow imaging, this study proposes a microbubble-based H-Scan ultrasound imaging technique. This technique utilizes high-order H-Scan to detect the Rayleigh scattering contributed by blood flow and microbubbles at certain concentrations. The detected results are then processed in the B channel using methods such as clutter filtering based on Casorati matrix singular value decomposition (Casorati-SVD). Compared with the control group without H-Scan, the signal-to-noise ratio (SNR) of the experimental group using the microbubble-based H-Scan ultrasound imaging technique was increased by 38.61% on average and the contrast signal-to-noise ratio (CNR) was increased by 39.5% on average. The improved image quality of microvascular flow imaging was visibly enhanced. This method demonstrates significant advantages in enhancing the sensitivity and accuracy of ultrasound blood flow imaging, indicating considerable potential for clinical applications.

Selective expression and significance of ACKR2 in lung aerocytes

BackgroundACKR2 is an atypical chemokine receptor that plays a significant role in regulating inflammation by binding to inflammatory CC chemokines and facilitating their degradation. Previous findings suggest that the genetic...

Evaluation of Real-Time Cardiovascular Flow MRI Using Compressed Sensing in a Phantom and in Patients With Valvular Disease or Arrhythmia.

Real-time (RT) phase contrast (PC) flow MRI can potentially be used to measure blood flow in arrhythmic patients. Undersampled RT PC has been combined with online compressed sensing (CS) reconstruction (CS RT) enabling clinical use. However, CS RT flow has not been validated in a clinical setting. Evaluate CS RT in phantom and patients. Prospective. Flow phantom (60 cycles/min: N = 10, 120 cycles/min: N = 12), sinus rhythm patients, no regurgitation (N = 20) or suspected aortic regurgitation (N = 10), arrhythmia patients (N = 10). 1.5 T, 2D gated PC, CS RT PC, RT cine with arrhythmia rejection. Phantom experiments tested the accuracy of CS RT cardiac output and peak flow rate at 60 and 120 cycles/min against gated PC. For sinus rhythm patients, cardiac output, peak flow rate, and regurgitation fraction in the ascending aorta and/or pulmonary artery were evaluated against gated PC. Cardiac output in patients with arrythmia was evaluated against RT cine with arrhythmia rejection. Bland Altman, correlation, Mann-Whitney test, Wilcoxon signed-rank test. Cardiac output bias ± SD for CS RT in the phantom was -0.0 ± 0.2 L/min (0.5 ± 3%, P = 0.76) at 60 cycles/min and 0.2 ± 0.3 L/min (4 ± 4%, P = 0.0016) at 120 cycles/min. Correspondingly, peak flow rate bias was -23 ± 6 mL/s (-7 ± 2%, P < 0.0001) and -73 ± 25 mL/s (-23 ± 4%, P < 0.0001). In patients, regurgitant fraction was -4 ± 0.5% (-23 ± 4%, P = 0.0025). Cardiac output bias in patients in sinus rhythm was -0.1 ± 0.5 L/min (-2 ± 10%, P = 0.99) (with regurgitation) and -0.3 ± 0.6 L/min (-5 ± 11%, P = 0.035) (without regurgitation). Peak flow rate bias was -60 ± 31 mL/s (-13 ± 6%, P < 0.0001) (with regurgitation) and -64 ± 32 mL/s (-16 ± 8%, P < 0.0001) (without regurgitation). Cardiac output bias was -0.4 ± 0.6 L/min (-9 ± 11%, P < 0.003) in arrhythmia patients. CS RT flow could potentially serve as a clinical tool for patients with or without valvular disease or arrhythmia, with accurate cardiac output and regurgitation fraction quantification. Accurate flow assessment is important in clinical evaluation of cardiac patients, but in the presence of irregular heart rhythm flow assessment is challenging. We have evaluated a new method using cardiac magnetic resonance imaging and real-time flow for blood flow assessment in cardiac patients. The method was tested against a reference method in a phantom flow model in low and high heart rates, and in cardiac patients with and without irregular heart rhythm and in different vessels. We found the cardiac magnetic resonance imaging real time flow method accurate and therefore promising for clinical implementation. 1 TECHNICAL EFFICACY: Stage 1.

Dual-modal radiomics ultrasound model to diagnose cervical lymph node metastases of differentiated thyroid carcinoma: a two-center study

ObjectivesTo establish and validate a dual-modal radiomics nomogram from grayscale ultrasound and color doppler flow imaging (CDFI) of cervical lymph nodes (LNs), aiming to improve the diagnostic accuracy of metastatic LNs in differentiated thyroid carcinoma (DTC).MethodsDTC patients with suspected cervical LNs in two medical centers were retrospectively enrolled. Pathological results were set as gold standard. We extracted radiomic characteristics from grayscale ultrasound and CDFI images, then applied lasso (least absolute shrinkage and selection operator) regression analysis to analyze radiomics features and calculate the rad-score. A nomogram based on rad-score, clinical data, and ultrasound signs was developed. The performance of the model was evaluated using AUC and calibration curve. We also assessed the model’s diagnostic ability in European Thyroid Association (ETA) indeterminate LNs.Results377 DTC patients and 726 LNs were enrolled. 37 radiomics features were determined and calculated as rad-score. The dual-modal radiomics model showed good calibration capabilities. The radiomics model displayed higher diagnostic ability than the traditional ultrasound model in the training set [0.871 (95% CI: 0.839–0.904) vs. 0.848 (95% CI: 0.812–0.884), p<0.01], internal test set [0.804 (95% CI: 0.741–0.867) vs. 0.803 (95% CI: 0.74–0.866), p = 0.696], and external validation cohort [0.939 (95% CI: 0.893–0.984) vs. 0.921 (95% CI: 0.857–0.985), p = 0.026]. The radiomics model could also significantly improve the detection rate of metastatic LNs in the ETA indeterminate LN category.ConclusionsThe dual-modal radiomics nomogram can improve the diagnostic accuracy of metastatic LNs of DTC, especially for LNs in ETA indeterminate classification.

Evaluation of Ceramic Membrane Filtration for Alternatives to Microplastics in Cosmetic Formulations Using FlowCam Analysis.

The rapid expansion of the cosmetics industry has significantly increased the adoption of alternative microplastics in response to increasingly stringent global environmental regulations. This study presents a comparative analysis of the treatment performance of silica powder and cornstarch-common alternatives for microplastics in cosmetics-using ceramic membrane filtration combined with flow imaging microscopy (FlowCam) to analyze particle behavior. Bench-scale crossflow filtration experiments were performed with commercially available alumina ceramic membranes. By analyzing high-resolution images from FlowCam, the transport and retention behaviors of the two microplastic alternatives were examined by comparing their morphological properties. Despite their similar particle sizes, the cornstarch demonstrated a higher removal efficiency (82%) than the silica (72%) in the ceramic membrane filtration due to its greater tendency to aggregate. This increased tendency for aggregation suggests that cornstarch may contribute to faster fouling, while the stability and uniformity of silica particles result in less fouling. The FlowCam analysis revealed that the cornstarch particles experienced a slight increase in circularity and compactness over time, likely due to physical swelling and aggregation, while the silica particles retained their shape and structural integrity. These findings highlight the impact of the morphological properties of alternative microplastics on their filtration behavior and fouling potential.

Detection and Characterization of Circulating Tumor Cells in Colorectal Cancer Patients via Epithelial-Mesenchymal Transition Markers.

Background/Objectives: Liquid biopsy methods have gained prominence as minimally invasive tools to improve cancer treatment outcomes. Circulating tumor cells (CTCs) offer valuable insights into both primary and metastatic lesions. However, validating the CTC test results requires confirmation that the detected cells originate from cancer tissue. While studies have identified CTCs in colorectal cancer (CRC) patients using molecular markers, simultaneous validation of their cancer tissue origin remains unexplored. Methods: This study introduces a simple approach to detect adenomatous polyposis coli (APC) gene abnormalities alongside established CTC markers using a molecular imaging flow cytometer (MI-FCM). Given that APC gene abnormalities occur in 60-70% of CRC patients, their detection serves as strong evidence of cancer origin. Results: Our method achieved 92% concordance with DNA sequence analysis of tumor-derived cells. In a proof-of-concept study using 5 mL of whole blood from CRC patients, we observed a high frequency of cells exhibiting APC abnormalities, cytokeratin (CK), and vimentin (Vim) expression. Extending the study to 80 CRC patients across pathological stages I-IV confirmed CK and Vim as valid CTC markers. Three distinct cell populations were identified in blood: CK+/Vim-, CK+/Vim+, and CK-/Vim+. CTC number and frequency increased progressively with cancer stage. Conclusions: This is the first report demonstrating CK and Vim as effective markers for direct CTC detection in CRC patients. Our findings provide evidence-based validation of CTC markers, offering new insights and advancing approaches for patient care.

Diagnostic Accuracy of Detective Flow Imaging Endoscopic Ultrasonography for Evaluating Blood Flow Within Mural Nodules of Intraductal Papillary Mucinous Neoplasms.

Background/Objectives: Detective flow imaging (DFI) endoscopic ultrasonography (EUS) can identify the microvascular flow imaging of a mural nodule (MN) in an intraductal papillary mucinous neoplasm (IPMN) without the use of contrast agents. This retrospective study evaluated the diagnostic accuracy of DFI-EUS and its ability to evaluate the blood flow of MNs in IPMNs. Methods: Between April 2021 and September 2023, 68 patients with MNs in IPMNs observed on EUS images were retrospectively analyzed. Both DFI-EUS and contrast-enhanced EUS (CE-EUS) were performed during the same session. Three expert endosonographers blinded to the patients' clinical data assessed the MN images obtained with CE-EUS and DFI-EUS. First, DFI-EUS images were evaluated using a predefined scoring system; thereafter, CE-EUS images were evaluated. The diagnostic capability of DFI-EUS to detect MN blood flow was assessed with CE-EUS as the gold standard. Secondary outcomes included inter-reader agreement, the correlation between MN size and detection rates, and the association between DFI blood flow signal patterns and malignancy of MNs in surgically resected cases. Results: CE-EUS showed a contrast effect in the MN in 24 cases. Among these, DFI-EUS detected blood flow signals in 20 cases; false-positive results were not observed. DFI-EUS demonstrated a sensitivity of 83%, specificity of 100%, and accuracy of 93% for detecting MN blood flow. Inter-reader agreement was substantial (kappa values, 0.6-0.8). The subgroup analysis revealed that all MNs ≥ 10 mm had detectable blood flow on DFI-EUS, whereas MNs < 10 mm had reduced detection rates (75%; 12/16 cases). No significant correlation between the DFI blood flow signal patterns and MN malignancy of resected cases was observed. Conclusions: DFI-EUS demonstrated high diagnostic accuracy for detecting MN blood flow. Because of its simplicity and cost-effectiveness, DFI-EUS could be an alternative to CE-EUS for patients with MNs inside IPMNs.

Accuracy of flow volume estimation in the dilated aorta using 4D flow MRI: a pulsatile phantom study.

Aortic dilatation is a severe pathology that increases the risk of rupture and its hemodynamics could be accurately assessed by using the 4D flow cardiovascular magnetic resonance (CMR) technique but flow assessment under complex flow patterns require validation. The aim of this work was to develop an in vitro system compatible with CMR to assess the accuracy of volume flow measurements in dilated aortas.&#xD;Approach. Two latex models, one with ascending and the other with abdominal aortic aneurysms were manufactured to ensure a constant and controlled net flow volume along the aortic length. A pneumatic piston driven by a stepper motor and controlled by an embedded system located in the control room modulated a pulsatile fluid flow using a pump with an elastic membrane placed in the magnet near the elastic models. All the visualization and measurement algorithms were integrated into a custom computer platform. 4D flow imaging was used to estimate the flow rate and volume through multiple aortic planes and compared to the reference assessed by weight method and to 2D flow measurements.&#xD;Main results. The errors of flow volume assessment using 4D flow remained within reasonable limits along the length of the aortic models. Mean differences in net flow volume from the reference were less than 2 ml (range -4 to 6 ml), corresponding to mean relative differences of less than 4% (range -8% to 11%). Averaged net, forward and backward flow volume estimations along the aortic length were similar using 2D and 4D flow measurements (p>0.05). Peak forward and backward flow rates increased in the dilated regions and were comparable to those observed in patients. &#xD;Significance. The accuracy of flow volume estimates in complex flow patterns, such as those observed in patients with aneurysms, was validated in vitro using 4D flow. &#xD.

The Impact of Targeted Therapies on Red Blood Cell Aggregation in Patients with Chronic Lymphocytic Leukemia Evaluated Using Software Image Flow Analysis.

Chronic lymphocytic leukemia (CLL), the most common type of leukemia, remains incurable with conventional therapy. Despite advances in therapies targeting Bruton's tyrosine kinase and anti-apoptotic protein BCL-2, little is known about their effect on red blood cell (RBC) aggregation in blood flow. In this study, we applied a microfluidic device and a newly developed Software Image Flow Analysis to assess the extent of RBC aggregation in CLL patients and to elucidate the hemorheological effects of the commonly applied therapeutics Obinutuzumab/Venetoclax and Ibrutinib. The results revealed that, in RBC samples from untreated CLL patients, complex 3D clusters of large RBC aggregates are formed, and their number is significantly increased compared to healthy control samples. The application of the Obinutuzumab/Venetoclax combination did not affect this aspect of RBCs' rheological behavior. In contrast, targeted therapy with Ibrutinib preserves the aggregation state of CLL RBCs to levels seen in healthy controls, demonstrating that Ibrutinib mitigates the alterations in the rheological properties of RBCs associated with CLL. Our findings highlight the alterations in RBC aggregation in CLL and the impact of different targeted therapies on RBCs' rheological properties, which is critical for predicting the potential complications and side effects of CLL treatments, particularly concerning blood flow dynamics.

Imaging Flow Cytometric Identification of Chromosomal Defects in Paediatric Acute Lymphoblastic Leukaemia.

Acute lymphoblastic leukaemia is the most common childhood malignancy that remains a leading cause of death in childhood. It may be characterised by multiple known recurrent genetic aberrations that inform prognosis, the most common being hyperdiploidy and t(12;21) ETV6::RUNX1. We aimed to assess the applicability of a new imaging flow cytometry methodology that incorporates cell morphology, immunophenotype, and fluorescence in situ hybridisation (FISH) to identify aneuploidy of chromosomes 4 and 21 and the translocation ETV6::RUNX1. We evaluated this new "immuno-flowFISH" platform on 39 cases of paediatric ALL of B-lineage known to have aneuploidy of chromosomes 4 and 21 and the translocation ETV6::RUNX1. After identifying the leukaemic population based on immunophenotype (i.e., expression of CD34, CD10, and CD19 antigens), we assessed for copy numbers of loci for the centromeres of chromosomes 4 and 21 and the ETV6 and RUNX1 regions using fluorophore-labelled DNA probes in more than 1000 cells per sample. Trisomy 4 and 21, tetrasomy 21, and translocations of ETV6::RUNX1, as well as gains and losses of ETV6 and RUNX1, could all be identified based on FISH spot counts and digital imagery. There was variability in clonal makeup in individual cases, suggesting the presence of sub-clones. Copy number alterations and translocations could be detected even when the cell population comprised less than 1% of cells and included cells with a mature B-cell phenotype, i.e., CD19-positive, lacking CD34 and CD10. In this proof-of-principle study of 39 cases, this sensitive and specific semi-automated high-throughput imaging flow cytometric immuno-flowFISH method has been able to show that alterations in ploidy and ETV6::RUNX1 could be detected in the 39 cases of paediatric ALL. This imaging flow cytometric FISH method has potential applications for diagnosis and monitoring disease and marrow regeneration (i.e., distinguishing residual ALL from regenerating haematogones) following chemotherapy.

Prognostic value of multi-PLD ASL radiomics in acute ischemic stroke.

Early prognosis prediction of acute ischemic stroke (AIS) can support clinicians in choosing personalized treatment plans. The aim of this study is to develop a machine learning (ML) model that uses multiple post-labeling delay times (multi-PLD) arterial spin labeling (ASL) radiomics features to achieve early and precise prediction of AIS prognosis. This study enrolled 102 AIS patients admitted between December 2020 and September 2024. Clinical data, such as age and baseline National Institutes of Health Stroke Scale (NIHSS) score, were collected. Radiomics features were extracted from cerebral blood flow (CBF) images acquired through multi-PLD ASL. Features were selected using least absolute shrinkage and selection operator regression, and three models were developed: a clinical model, a CBF radiomics model, and a combined model, employing eight ML algorithms. Model performance was assessed using receiver operating characteristic curves and decision curve analysis (DCA). Shapley Additive exPlanations was applied to interpret feature contributions. The combined model of extreme gradient boosting demonstrated superior predictive performance, achieving an area under the curve (AUC) of 0.876. Statistical analysis using the DeLong test revealed its significant outperformance compared to both the clinical model (AUC = 0.658, p < 0.001) and the CBF radiomics model (AUC = 0.755, p = 0.002). The robustness of all models was confirmed through permutation testing. Furthermore, DCA underscored the clinical utility of the combined model. The prognostic prediction of AIS was notably influenced by the baseline NIHSS score, age, as well as texture and shape features of CBF. The integration of clinical data and multi-PLD ASL radiomics features in a model offers a secure and dependable approach for predicting the prognosis of AIS, particularly beneficial for patients with contraindications to contrast agents. This model aids clinicians in devising individualized treatment plans, ultimately enhancing patient prognosis.