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1.
Methods Mol Biol ; 2857: 45-59, 2025.
Article in English | MEDLINE | ID: mdl-39348054

ABSTRACT

Flow cytometry serves as a crucial tool in immunology, allowing for the detailed analysis of immune cell populations. γδ T cells, a subset of T cells, play pivotal roles in immune surveillance and immune aging. Assessing the phenotype and functional capabilities of γδ T cells isolated from whole blood or tissue within the context of human aging yields invaluable insights into the dynamic changes affecting immune function, tissue homeostasis, susceptibility to infections, and inflammatory responses.


Subject(s)
Aging , Flow Cytometry , Immunophenotyping , Receptors, Antigen, T-Cell, gamma-delta , Humans , Immunophenotyping/methods , Aging/immunology , Flow Cytometry/methods , Receptors, Antigen, T-Cell, gamma-delta/metabolism , Receptors, Antigen, T-Cell, gamma-delta/immunology , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism , T-Lymphocytes/immunology
2.
Methods Mol Biol ; 2857: 137-146, 2025.
Article in English | MEDLINE | ID: mdl-39348062

ABSTRACT

Extracellular vesicles (EVs) are lipid-bound particles produced by a wide variety of cells from different biological species. EVs can carry molecules, such as nucleic acids and metabolites, and are involved in cell functioning, communication, and signaling. Recent literature reported that pathogenic or commensal yeast strains can produce EVs targeting the host's immune system and exerting immunomodulatory actions. In humans, yeast EVs can be endocytosed by dendritic cells (DCs), characterized by phagocyting and migrating capabilities with the role of capturing antigens to present to T lymphocytes, triggering the immune response. Physiological or disease-associated immunosenescence impairs both DC functionality and gut microbiota; thus investigating the interaction between commensal microorganisms and the host's immune system would help elucidate the impact of aging on the immune system-microbiota interplay. We hereby present a protocol for the incubation of in vitro-generated human monocyte-derived DCs with EVs purified from different yeast strains isolated from fermented milk. The protocol includes flow cytometry analysis on DC activation markers and endocytosis assay.


Subject(s)
Dendritic Cells , Extracellular Vesicles , Monocytes , Humans , Dendritic Cells/metabolism , Dendritic Cells/immunology , Extracellular Vesicles/metabolism , Extracellular Vesicles/immunology , Monocytes/metabolism , Monocytes/immunology , Monocytes/microbiology , Flow Cytometry/methods , Endocytosis , Yeasts/metabolism , Saccharomyces cerevisiae/metabolism , Cells, Cultured
3.
Methods Mol Biol ; 2857: 15-31, 2025.
Article in English | MEDLINE | ID: mdl-39348052

ABSTRACT

B cells are crucial components of the immune system, responsible for producing specific antibodies in response to infections and vaccines. Despite their uniform appearance, B cells display diverse surface molecules and functional properties, which are not yet fully understood. Apart from antibody production, B cells also play roles in antigen presentation and cytokine secretion, essential for initiating T-cell immune responses. Their significance as disease biomarkers and therapeutic targets has led to increased research focus. However, the lack of standardized protocols for B-cell identification and the variability in defining B-lymphocyte subpopulations pose some challenges. This paper proposes a B-cell identification panel throughout the evaluation of previous cytometry panels and nomenclature heterogeneity for B-cell subpopulations. Major subpopulations recognized in human peripheral blood include transitional, naive, switched memory, unswitched memory, double negative, and plasmablasts, characterized based on their functional and phenotypic features. We present a standardized flow cytometry protocol utilizing surface phenotypic markers (CD3, CD19, IgD, CD27, CD38, and CD24) to differentiate and analyze B-cell subpopulations. This practical and cost-effective panel can be used in various research and laboratory settings. The challenges of standardizing names and markers for classifying B-lymphocyte subpopulations are discussed, along with protocols utilizing multiple markers and gating strategies, allied with the importance of considering viability markers. In summary, this standardized protocol and panel provide a comprehensive approach to identifying B-cell subpopulations to enhance the reproducibility and comparability of B-cell subpopulation studies.


Subject(s)
B-Lymphocyte Subsets , Flow Cytometry , Immunophenotyping , Humans , Flow Cytometry/methods , Immunophenotyping/methods , B-Lymphocyte Subsets/immunology , B-Lymphocyte Subsets/metabolism , B-Lymphocyte Subsets/cytology , B-Lymphocytes/immunology , B-Lymphocytes/cytology , B-Lymphocytes/metabolism , Biomarkers , Phenotype , Antigens, CD/immunology , Antigens, CD/metabolism , Cost-Benefit Analysis
4.
Methods Mol Biol ; 2857: 169-180, 2025.
Article in English | MEDLINE | ID: mdl-39348065

ABSTRACT

Acute skeletal muscle injury initiates a process of necrosis, debris clearance, and ultimately tissue regeneration via myogenesis. While skeletal muscle stem cells (MuSCs) are responsible for populating the proliferative myogenic progenitor pool to fuel muscle repair, recruited and resident immune cells have a central role in the regulation of muscle regeneration via the execution of phagocytosis and release of soluble factors that act directly on MuSCs to regulate myogenic differentiation. Therefore, the timing of MuSC proliferation and differentiation is closely linked to the populations and behaviors of immune cells present within skeletal muscle. This has important implications for aging and muscle repair, as systemic changes in immune system function contribute to a decline in muscle regenerative capacity. Here, we present adapted protocols for the isolation of mononuclear cells from skeletal muscles for the quantification of immune cell populations using flow cytometry. We also describe a cardiotoxin skeletal muscle injury protocol and detail the expected outcomes including immune cell infiltration to the injured sites and formation of new myocytes. As immune cell function is substantially influenced by aging, we extend these approaches and outcomes to aged mice.


Subject(s)
Aging , Disease Models, Animal , Muscle, Skeletal , Regeneration , Animals , Mice , Muscle, Skeletal/injuries , Muscle, Skeletal/metabolism , Aging/physiology , Muscle Development , Flow Cytometry/methods , Cell Differentiation , Cell Proliferation
5.
Methods Mol Biol ; 2855: 373-385, 2025.
Article in English | MEDLINE | ID: mdl-39354319

ABSTRACT

Cardiolipins (CL) are special lipids in many respects. First of all, CL are composed of four fatty acids linked by two phosphatidic acids, which provide CL a unique molecular structure. Secondly, in eukaryotic cells they are specific to a single organelle, mitochondria, where they are also synthetized. CL are one of the most abundant lipid classes in mitochondria, mainly localized in the inner membrane. They are key determinants of mitochondrial health and homeostasis by modulating membrane integrity and fluidity, mitochondrial shapes, and metabolic pathways. Disturbances in mitochondrial CL composition can lead to tissue malfunction and diseases. It is therefore important to develop analytical tools to study the mitochondrial lipidome, and more particularly the CL. The method described here allows the quantification of cardiolipins at the sum composition level in isolated mitochondria or in liver tissue by flow injection analysis coupled to differential mobility spectrometry (FIA-DMS), also known as DMS-based shotgun lipidomics.


Subject(s)
Cardiolipins , Lipidomics , Cardiolipins/analysis , Cardiolipins/metabolism , Lipidomics/methods , Animals , Mitochondria/metabolism , Mass Spectrometry/methods , Liver/metabolism , Liver/chemistry
6.
Clin Chim Acta ; 564: 119927, 2025 Jan 01.
Article in English | MEDLINE | ID: mdl-39153656

ABSTRACT

BACKGROUND: Helicobacter pylori (H. pylori) infects over 50% of the global population and is a significant risk factor for gastric cancer. The pathogenicity of H. pylori is primarily attributed to virulence factors such as vacA. Timely and accurate identification, along with genotyping of H. pylori virulence genes, are essential for effective clinical management and controlling its prevalence. METHODS: In this study, we developed a dual-target RAA-LFD assay for the rapid, visual detection of H. pylori genes (16s rRNA, ureA, vacA m1/m2), using recombinase aided amplification (RAA) combined with lateral flow dipstick (LFD) methods. Both 16s rRNA and ureA were selected as identification genes to ensure reliable detection accuracy. RESULTS: A RAA-LFD assay was developed to achieve dual-target amplification at a stable 37 °C within 20 min, followed by visualization using the lateral flow dipstick (LFD). The whole process, from amplification to results, took less than 30 min. The 95 % limit of detection (LOD) for 16 s rRNA and ureA, vacA m1, vacA m2 were determined as 3.8 × 10-2 ng/µL, 5.8 × 10-2 ng/µL and 1.4 × 10-2 ng/µL, respectively. No cross-reaction was observed in the detection of common pathogens including Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus subtilis, showing the assay's high specificity. In the evaluation of the clinical performance of the RAA-LFD assay. A total of 44 gastric juice samples were analyzed, immunofluorescence staining (IFS) and quantitative polymerase chain reaction (qPCR) were used as reference methods. The RAA-LFD results for the 16s rRNA and ureA genes showed complete agreement with qPCR findings, accurately identifying H. pylori infection as confirmed by IFS in 10 out of the 44 patients. Furthermore, the assay successfully genotyped vacA m1/m2 among the positive samples, showing complete agreement with qPCR results and achieving a kappa (κ) value of 1.00. CONCLUSION: The dual-target RAA-LFD assay developed in this study provides a rapid and reliable method for detecting and genotyping H. pylori within 30 min, minimizing dependency on sophisticated laboratory equipment and specialized personnel. Clinical validation confirms its efficacy as a promising tool for effectively control of its prevalence and aiding in the precise treatment of H. pylori-associated diseases.


Subject(s)
Bacterial Proteins , Helicobacter pylori , Helicobacter pylori/genetics , Helicobacter pylori/isolation & purification , Bacterial Proteins/genetics , Humans , RNA, Ribosomal, 16S/genetics , Helicobacter Infections/diagnosis , Helicobacter Infections/microbiology , Nucleic Acid Amplification Techniques/methods
7.
J Clin Exp Hepatol ; 15(1): 102403, 2025.
Article in English | MEDLINE | ID: mdl-39296664

ABSTRACT

Background/Aims: In this study, we aim to develop a model for predicting gastroesophageal varices (GEV) bleeding in patients with chronic hepatitis B (CHB) by utilizing hemodynamic parameters obtained through four-dimensional flow MRI (4D flow MRI). Methods: This study conducted a prospective enrollment of CHB patients suspected of GEV from October 2021 to May 2022. The severity of varices and bleeding risk were evaluated using clinical findings and upper gastrointestinal endoscopy, and patients were classified into high-risk and non-high-risk groups. The study utilized serological examination, ultrasonographic examination, and 4D flow MRI. Relevant parameters were selected through univariate and multivariate analyses, and a prediction model was established using binary logistic regression analysis. The model was combined with the Baveno Ⅵ/Ⅶ and Expanded Baveno Ⅵ/Ⅶ criteria to evaluate diagnostic efficacy and the risk of avoiding endoscopic examination. Results: A total of 40 CHB patients were enrolled and categorized into the high-risk group (n = 15) and the non-high-risk group (n = 25). The spleen diameter and regurgitant fraction (R%) were independent predictors of variceal bleeding and a predictive model was established. The combination of this prediction model and the Baveno Ⅵ/Ⅶ criteria achieved high diagnostic efficiency, enabling 45.00% (18/40) of patients to be exempted from the unnecessary endoscopic procedure and the high-risk misclassification rate (0%) was less than 5%. Conclusion: The prediction model generated by 4D flow MRI has the potential to assess the likelihood of varices and can be supplemented by the Baveno VI/VII criteria to improve diagnostic accuracy in CHB patients.

8.
Methods Mol Biol ; 2852: 105-122, 2025.
Article in English | MEDLINE | ID: mdl-39235739

ABSTRACT

In food industry, Listeria monocytogenes contamination can occur accidentally despite the quality control of raw materials and factory. Decontamination processes or inhibitory effects of ingredients/additives in food products are set up to ensure compliance with hygiene and microbiological criteria. These actions represent stresses for the pathogenic agent, causing fluctuations in its physiological states. Moreover, during these environmental stresses, Listeria monocytogenes can enter in a viable but nonculturable (VBNC) state which is not detected by plate counting but by flow cytometry. This technique coupled with cell staining by fluorescent dyes offers the possibility to assess different physiological states based on different cellular parameters: enzymatic activity, transmembrane integrity, membrane potential, and respiratory activity. In this chapter, we present a method to assess the viability of foodborne pathogens using a double-staining principle based on the assessment of membrane integrity and intracellular esterase activity.


Subject(s)
Flow Cytometry , Listeria monocytogenes , Microbial Viability , Listeria monocytogenes/growth & development , Listeria monocytogenes/physiology , Flow Cytometry/methods , Food Microbiology/methods , Fluorescent Dyes/chemistry , Staining and Labeling/methods , Cell Membrane/metabolism
9.
Clin Chim Acta ; 564: 119945, 2025 Jan 01.
Article in English | MEDLINE | ID: mdl-39209245

ABSTRACT

Acute myeloid leukemia (AML) is a common type of acute leukemia (AL), belonging to malignant tumors of the hematopoietic system with the characteristics of rapid disease development, control with extreme difficulties, easy recurrence, poor prognosis, and incidence rate increasing with age. The traditionally diagnostic standard of French American British (FAB), being based on the morphological examination with high human subjectivity, can no longer meet the demand of clinical diagnosis and treatment of AML. Requirements of objective accuracy and low-dose sample, have become the indispensable method for AML diagnosis and monitoring prognosis. Flow cytometry is a modern technology that can quickly and accurately detect the series, antigen distribution, differentiation stage of AML cells, minimal residual lesions after AML therapy, so as to provide the great significance in guiding clinical diagnosis, hierarchical treatment, and prognosis judgement. This article will systematically elaborate on the application of flow cytometry in the diagnosis and classification of AML, and the detection of minimal residual lesions, thereby providing reference significance for dynamic monitoring and prognostic observation of AML with different immune subtypes of FAB.


Subject(s)
Flow Cytometry , Leukemia, Myeloid, Acute , Neoplasm, Residual , Humans , Leukemia, Myeloid, Acute/diagnosis , Neoplasm, Residual/diagnosis
10.
Methods Mol Biol ; 2857: 79-87, 2025.
Article in English | MEDLINE | ID: mdl-39348056

ABSTRACT

Rheumatoid arthritis (RA) is linked to various signs of advanced aging, such as premature immunosenescence which occurs due to decline in regenerative ability of T cells. RA T cells develop a unique aggressive inflammatory senescent phenotype with an imbalance of Th17/T regulatory (Treg) cell homeostasis and presence of CD28- T cells. The phenotypic analysis and characterization of T cell subsets become necessary to ascertain if any functional deficiencies exist within with the help of transcription factor (TF) analysis. These subset-specific TFs dictate the functional characteristics of T-cell populations, leading to the production of distinct effector cytokines and functions. Examining the expression, activity, regulation, and genetic sequence of TFs not only aids researchers in determining their importance in disease processes but also aids in immunological monitoring of patients enrolled in clinical trials, particularly in evaluating various T-cell subsets [Th17 (CD3+CD4+IL17+RORγt+) cells and T regulatory (Treg) (CD3+CD4+CD25+CD127-FOXP3+) cells], markers of T-cell aging [aged Th17 cells (CD3+CD4+IL17+RORγt+CD28-), and aged Treg cells (CD3+CD4+CD25+CD127-FOXP3+CD28-)]. In this context, we propose and outline the protocols for assessing the expression of TFs in aged Th17 and Treg cells, highlighting the crucial aspects of this cytometric approach.


Subject(s)
Arthritis, Rheumatoid , Immunosenescence , T-Lymphocytes, Regulatory , Transcription Factors , Humans , Arthritis, Rheumatoid/immunology , Arthritis, Rheumatoid/metabolism , Arthritis, Rheumatoid/genetics , Transcription Factors/metabolism , Transcription Factors/genetics , T-Lymphocytes, Regulatory/immunology , T-Lymphocytes, Regulatory/metabolism , Th17 Cells/immunology , Th17 Cells/metabolism , Flow Cytometry/methods , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism , Biomarkers
11.
Front Cardiovasc Med ; 11: 1398114, 2024.
Article in English | MEDLINE | ID: mdl-39355352

ABSTRACT

Aims: Patients with severe aortic stenosis (AS), low transvalvular flow (LF) and low gradient (LG) with normal ejection fraction (EF)-are referred to as paradoxical LF-LG AS (PLF-LG). PLF-LG patients develop more advanced heart failure symptoms and have a worse prognosis than patients with normal EF and high-gradient AS (NEF-HG). Despite its clinical relevance, the mechanisms underlying PLF-LG are still poorly understood. Methods: Left ventricular (LV) myocardial biopsies of PLF-LG (n = 5) and NEF-HG patients (n = 6), obtained during transcatheter aortic valve implantation, were analyzed by LC-MS/MS after sequential extraction of cellular and extracellular matrix (ECM) proteins using a three-step extraction method. Proteomic data are available via ProteomeXchange with identifier PXD055391. Results: 73 cellular proteins were differentially abundant between the 2 groups. Among these, a network of proteins related to muscle contraction and arrhythmogenic cardiomyopathy (e.g., cTnI, FKBP1A and CACNA2D1) was found in PLF-LG. Extracellularly, upregulated proteins in PLF-LG were related to ATP synthesis and oxidative phosphorylation (e.g., ATP5PF, COX5B and UQCRB). Interestingly, we observed a 1.3-fold increase in cyclophilin A (CyPA), proinflammatory cytokine, in the extracellular extracts of PLF-LG AS patients (p < 0.05). Consistently, immunohistochemical analysis confirmed its extracellular localization in PLF-LG AS LV sections along with an increase in its receptor, CD147, compared to the NEF-HG AS patients. Levels of core ECM proteins, namely collagens and proteoglycans, were comparable between groups. Conclusion: Our study pinpointed novel candidates and processes with potential relevance in the pathophysiology of PLF-LG. The role of CyPA in particular warrants further investigation.

12.
World J Radiol ; 16(9): 429-438, 2024 Sep 28.
Article in English | MEDLINE | ID: mdl-39355390

ABSTRACT

BACKGROUND: Global and regional cerebral blood flow (CBF) changes in patients with unilateral internal carotid artery occlusion (ICAO) are unclear when the dual post-labeling delays (PLD) arterial spin labeling (ASL) magnetic resonance imaging (MRI) technique is used. Manual delineation of regions of interest for CBF measurement is time-consuming and laborious. AIM: To assess global and regional CBF changes in patients with unilateral ICAO with the ASL-MRI perfusion technique. METHODS: Twenty hospitalized patients with ICAO and sex- and age-matched controls were included in the study. Regional CBF was measured by Dr. Brain's ASL software. The present study evaluated differences in global, middle cerebral artery (MCA) territory, anterior cerebral artery territory, and Alberta Stroke Program Early Computed Tomography Score (ASPECTS) regions (including the caudate nucleus, lentiform nucleus, insula ribbon, internal capsule, and M1-M6) and brain lobes (including frontal, parietal, temporal, and insular lobes) between ICAO patients and controls at PLD 1.5 s and PLD 2.5 s. RESULTS: When comparing CBF between ICAO patients and controls, the global CBF in ICAO patients was lower at both PLD 1.5 s and PLD 2.5 s; the CBF on the occluded side was lower in 15 brain regions at PLD 1.5 s, and it was lower in 9 brain regions at PLD 2.5 s; the CBF in the contralateral hemisphere was lower in the caudate nucleus and internal capsule at PLD 1.5 s and in M6 at PLD 2.5 s. The global CBF in ICAO patients was lower at PLD 1.5 s than at PLD 2.5 s. The ipsilateral CBF at PLD 1.5 s was lower than that at PLD 2.5 s in 15 regions, whereas the contralateral CBF was lower at PLD 1.5 s than at PLD 2.5 s in 12 regions. The ipsilateral CBF was lower than the contralateral CBF in 15 regions at PLD 1.5 s, and in M6 at PLD 2.5 s. CONCLUSION: Unilateral ICAO results in hypoperfusion in the global and MCA territories, especially in the ASPECTS area. Dual PLD settings prove more suitable for accurate CBF quantification in ICAO.

13.
Front Aging Neurosci ; 16: 1457675, 2024.
Article in English | MEDLINE | ID: mdl-39355539

ABSTRACT

Background: The mechanism of cognitive impairment in hemodialysis patients is multifactorial. The relationship between cerebral blood flow and the decline of cognitive function is poorly understood. Objective: To investigate the association between cerebral blood flow variation and decline of cognitive function in older patients undergoing hemodialysis. Methods: In this prospective observational cohort study of 121 older patients undergoing hemodialysis, we used transcranial Doppler ultrasound (TCD) to measure cerebral arterial mean flow velocity (MFV) throughout dialysis, assessed cognitive function at baseline and 12-month follow-up, and then analyzed associations between MFV and changes on cognitive scores. Results: TCD recordings demonstrated a significant reduction in MFV throughout dialysis, which were significantly correlated with cumulative ultrafiltration volume (rho 0.356, p < 0.001), ΔSBP (rho 0.251, p = 0.005), and ΔMAP (rho 0.194, p = 0.032). Compared with the baseline assessments, cognitive scores of participants at the 12-month follow-up were significantly worsened in global cognition (MOCA), some tests of memory (CFT-memory), executive function (TMT-B, SCWT-C, and SCWT-T), attention/processing speed (SDMT), and visuospatial function (CFT-copy) (p < 0.05). The worsening scores in global cognition (MOCA) (ß = 0.066, 95% CI 0.018-0.113, p = 0.007) and some tests of memory (AVLT5) (ß = 0.050, 95% CI 0.004-0.097, p = 0.035) and executive function (TMT-B, SCWT-C, SCWT-T) (ß = 1.955, 95% CI 0.457-3.453, p = 0.011; ß = 0.298, 95% CI 0.112-0.484, p = 0.002 and ß = 1.371, 95% CI 0.429-2.303, p = 0.004, respectively) were significantly associated with the reduction of MFV. Conclusion: Hemodialysis may significantly reduce cerebral blood flow in older patients; Repetitive intradialytic decreases in CBF may be one of the mechanisms underlying the decline of cognitive function. Clinical trial registration: https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S000C5B5&selectaction=Edit&uid=U0003QEL&ts=4&cx=-djoi2.

14.
Rev Cardiovasc Med ; 25(9): 329, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39355580

ABSTRACT

Background: A fractional flow reserve (FFR)-fixed-SYNTAX score could decrease the number of high-risk patients. This study explored the prognostic value of non-invasive quantitative flow ratio (QFR)-fixed-SYNTAX I/II scores in multivessel disease patients. Methods: This was a single-center, small-sample, observational study. Multivessel coronary disease patients were enrolled and finished a 1-year follow-up. SYNTAX scores I/II and functional SYNTAX scores I/II based on QFR (cut-off value of 0.85) were calculated for all patients. The composite occurrence of cardiac deaths, any myocardial infarction, or ischemia-driven revascularization were analyzed using a different score system. Results: A total of 160 patients were stratified into risk groups based on a different scoring system. FSS (functional SYNTAX score) and FSSII (functional SYNTAX score II) reduce the radio of high-risk major adverse cardiovascular events (MACEs), transforming the patients from high-risk to medium- and low-risk. Furthermore, FSSII (hazard ratio (HR): 1.069, 95% CI: 1.025-1.115, p = 0.002) showed a better relationship with MACEs than the other score systems. After recalculating SSII, the survival-free ratio stratified by FSSII decreased from 38.46% to 27.27% in the high-risk group and increased from 84.09% to 86.05% in the low-risk group. Conclusions: FSS or FSSII could decrease the number of high-risk patients compared to SYNTAX score (SS) and FSS. SYNTAX II score (SSII) and FSSII showed a better predictive ability than other scoring systems for under-risk stratification.

15.
ACS Sens ; 2024 Oct 02.
Article in English | MEDLINE | ID: mdl-39356828

ABSTRACT

The early diagnosis of cancer in a point-of-need manner is of great significance, yet it remains challenging to achieve the necessary sensitivity and speed. Traditional lateral flow immunoassay (LFIA) methods are limited in accuracy and quantification, restricting their suitability for home-based applications. Thus, we explored a new and user-friendly electrochemical LFIA (e-LFIA) test strip to detect α-fetoprotein (AFP), a diagnostic marker for liver cancer. The specific electrochemical immunoprobe utilized in this e-LFIA test strip is characterized by significant signal boosting, resulted from the loading Ag shell into a gold nanoparticle (AuNP)-coated dendritic mesoporous silica nanoscaffold (DMSN). Leveraging the distinct electrochemical characteristics of Ag anodic stripping and the high volume-to-surface area ratio of DMSNs, the developed DMSNs/AuNPs@Ag-based e-LFIA test strip is capable of detecting AFP at a low concentration of 0.85 ng/mL within a rapid 20 min timespan, both of these values are smaller than those in current clinical testing. Furthermore, we utilized homemade screen-printed electrodes in this sensing prototype and demonstrated the high versatility and reliability of this e-LFIA device. We envision that this DMSNs/AuNPs@Ag-based e-LFIA holds substantial potential for the early diagnosis of liver cancer and household health monitoring.

16.
J Chromatogr A ; 1736: 465393, 2024 Sep 22.
Article in English | MEDLINE | ID: mdl-39357173

ABSTRACT

Three-dimensional (3D) cancer models, such as multicellular tumor spheroids (MCTS), are biological supports used for research in oncology, drug development and nanotoxicity assays. However, due to various analytical and biological challenges, the main recurring problem faced when developing this type of 3D model is the lack of reproducibility. When using a 3D support to assess the effect of biologics, small molecules or nanoparticles, it is essential that the support remains constant over time and multiples productions. This constancy ensures that any effect observed following molecule exposure can be attributed to the molecule itself and not to the heterogeneous properties of the 3D support. In this study, we address these analytical challenges by evaluating for the first time the 3D culture of a sub-population of cancer stem cells (CSCs) from a glioblastoma cancer cell line (U87-MG), produced by a SdFFF (sedimentation field-flow fractionation) cell sorting, in a supramolecular hydrogel composed of single, well-defined molecule (bis-amide bola amphiphile 0.25% w/v) with a stiffness of 0.4 kPa. CSCs were chosen for their ability of self-renewal and multipotency that allow them to generate fully-grown tumors from a small number of cells. The results demonstrate that CSCs cultured in the hydrogel formed spheroids with a mean diameter of 336.67 ± 38.70 µm by Day 35, indicating reproducible growth kinetics. This uniformity is in contrast with spheroids derived from unsorted cells, which displayed a more heterogeneous growth pattern, with a mean diameter of 203.20 ± 102.93 µm by Day 35. Statistical analysis using an unpaired t-test with unequal variances confirmed that this difference in spheroid size is significant, with a p-value of 0.0417 (p < 0.05). These findings demonstrate that CSC-derived spheroids, when cultured in a well-defined hydrogel, exhibit highly reproducible growth patterns compared to spheroids derived from unsorted cells, making them a more reliable 3D model for biological research and drug testing applications.

17.
Clin Neurol Neurosurg ; 246: 108555, 2024 Sep 22.
Article in English | MEDLINE | ID: mdl-39357321

ABSTRACT

BACKGROUND: Cranial nerve (CN) palsies are rare presenting symptoms of intracranial aneurysms. Our objectives were to report our institutional outcomes and study-level meta-analysis summarizing rates of improvement and identifying factors associated with recovery from CN symptoms after flow diversion. METHODS: We conducted a retrospective review of our institutional database for patients with intracranial aneurysms presenting with CN palsies who underwent treatment with flow diversion between 2015 and 2023. Systematic review of the literature was performed using Medline, EMBASE, Cochrane, as well as manual citation searches. Random effects meta-analysis was used. RESULTS: Thirteen of 136 studies were included in the meta-analysis and were combined with our institutional data. The pooled rate of improvement in any CN palsies following flow diversion was 71 % (95 %CI, 60 %-82 %, n=322). Patients presenting with CN II deficits were less likely to improve following treatment compared to other CN deficits (pooled OR [pOR] 0.32, 95 %CI, 0.16-0.63, n=224). The pooled rate of clinical improvement was 53 % in CNII deficits (95 %CI, 42 %-65 %, n=80) and 80 % in other CN deficits (95 %CI, 71 %-88 %, n=106). An increased rate of improvement was associated with acute intervention (pOR 9.12, 95 % CI, 2.26-36.73, n = 71) and radiographic aneurysm occlusion (pOR 5.29, 95 %CI, 1.66-16.90, n=118). CONCLUSIONS: Flow diversion improves CN palsy outcomes in patients with symptomatic intracranial aneurysms. The lower rate of improvement in visual acuity compared to other CN deficits may point to a different mechanism of injury or potential recoverability in these patients.

18.
Int J Surg Case Rep ; 124: 110357, 2024 Sep 27.
Article in English | MEDLINE | ID: mdl-39357475

ABSTRACT

INTRODUCTION: Whether aberrant arteries in pulmonary sequestration should be divided at the bifurcation or at the periphery remains a subject of debate (Kodama et al., 2016). Due to the risk of postoperative aneurysm formation followed by fetal bleeding (Rubin et al., 1994), it is thought that the aberrant artery should be divided at the bifurcation in cases of pulmonary sequestration. PRESENTATION OF CASE: A 35-year-old woman was referred to our hospital with continuous cough. Enhanced computed tomography (CT) subsequently revealed an infiltrative appearance in the right lower lobe of the lung, with an aberrant artery that originated from the left gastric artery and flowed into the right lower lobe of the lung. The aberrant artery was divided at the pulmonary ligament level; right lower lobectomy was performed. Enhanced CT performed four months postoperatively revealed that the residual region of the aberrant artery was spontaneously occluded. DISCUSSION: Although aberrant arteries are generally resected at their bifurcations, the present case suggests that dividing at the bifurcation might not be necessary. Alternatively, there may be characteristics that make resection at the bifurcation unnecessary. CONCLUSION: Herein, we present a case wherein the residual aberrant artery was spontaneously occluded following resection of pulmonary sequestration.

19.
Cell Metab ; 36(10): 2173-2189, 2024 Oct 01.
Article in English | MEDLINE | ID: mdl-39357509

ABSTRACT

Readily available nutrient-rich foods exploit our inherent drive to overconsume, creating an environment of overnutrition. This transformative setting has led to persistent health issues, such as obesity and metabolic syndrome. The development of glucagon-like peptide-1 receptor (GLP-1R) agonists reveals our ability to pharmacologically manage weight and address metabolic conditions. Obesity is directly linked to chronic low-grade inflammation, connecting our metabolic environment to neurodegenerative diseases. GLP-1R agonism in curbing obesity, achieved by impacting appetite and addressing associated metabolic defects, is revealing additional benefits extending beyond weight loss. Whether GLP-1R agonism directly impacts brain health or does so indirectly through improved metabolic health remains to be elucidated. In exploring the intricate connection between obesity and neurological conditions, recent literature suggests that GLP-1R agonism may have the capacity to shape the neurovascular landscape. Thus, GLP-1R agonism emerges as a promising strategy for addressing the complex interplay between metabolic health and cognitive well-being.


Subject(s)
Glucagon-Like Peptide 1 , Glucagon-Like Peptide-1 Receptor , Humans , Glucagon-Like Peptide 1/metabolism , Animals , Glucagon-Like Peptide-1 Receptor/agonists , Glucagon-Like Peptide-1 Receptor/metabolism , Obesity/metabolism , Brain/metabolism
20.
Phys Med Biol ; 2024 Oct 02.
Article in English | MEDLINE | ID: mdl-39357539

ABSTRACT

OBJECTIVE: Blood flow sensitivity is a crucial metric for appraising the effectiveness of color Doppler flow imaging (CDFI). Color Doppler velocity maps based on classic autocorrelation techniques are widely used in clinical practice. However, these techniques often produce twinkling artifacts in noisy regions due to the inherent randomness of noise phases. To mitigate artifacts and improve image quality, Power Mask (PoM) technology becomes imperative. Nevertheless, PoM technology unintentionally filters out small flow signals that have similar power and frequency characteristics to noise signals, thereby reducing the imaging system's sensitivity to flow. Approach: To address this issue, a novel Flow Recycling Algorithm (FRA) based on phase anomaly is introduced in this study. This algorithm, excavating small flow signals from noise, aims to enhance the small flow signals with low-velocity by the phase characteristics of the color Doppler flow information. Main results: Experiments in multi-organ imaging have shown that the FRA-CDFI approach is more effective in suppressing twinkling artifacts in noisy regions, preserving intricate small flow signals, and markedly improving small blood flow sensitivity. This novel approach provides adequate technical support for clinical ultrasound imaging of organs with dense small blood vessels, such as the brain, kidneys, liver, and more. Significance: As a novel post-processing method, FRA-CDFI holds significant potential for future deployment in clinical high-frame-rate ultrasound imaging devices.

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