Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 11.787
Filter
1.
Nutrients ; 16(17)2024 Sep 04.
Article in English | MEDLINE | ID: mdl-39275304

ABSTRACT

BACKGROUND: An appropriate diet is the basis for the treatment of type 2 diabetes (T2DM). However, there are no strict recommendations regarding the content of micronutrients and their modifications in the presence of chronic kidney disease (CKD). Therefore, we decided to investigate whether T2DM patients, including those with CKD, have different levels of chromium, nickel, cobalt, magnesium, and zinc in various blood elements compared to healthy individuals. METHODS: We divided our subjects into three groups: the control group (individuals without T2DM and proper renal function), those with T2DM and proper renal function, and those with T2DM and GFR < 60 mL/min/1.73 m2. RESULTS: We observed higher levels of chromium in all materials examined in patients with T2DM and impaired renal function. Both study groups found higher levels of nickel in samples of whole blood and red blood cells. Patients with T2DM and proper renal function had higher levels of serum manganese. Both study groups had lower levels of serum zinc. We observed higher levels of chromium in all materials examined in patients with T2DM and impaired renal function. Both study groups found higher levels of nickel in samples of whole blood and red blood cells. Patients with T2DM and proper renal function had higher levels of serum manganese. Both study groups had lower levels of serum zinc. CONCLUSIONS: In order to ensure effective care for patients with T2DM, it is necessary to improve the standard diet, including the content of micronutrients and their modification in patients with concomitant CKD.


Subject(s)
Diabetes Mellitus, Type 2 , Renal Insufficiency, Chronic , Trace Elements , Humans , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/complications , Trace Elements/blood , Male , Female , Middle Aged , Renal Insufficiency, Chronic/blood , Aged , Nickel/blood , Chromium/blood , Adult , Glomerular Filtration Rate , Zinc/blood , Magnesium/blood , Blood Cells , Case-Control Studies
2.
Cells ; 13(17)2024 Aug 30.
Article in English | MEDLINE | ID: mdl-39273028

ABSTRACT

Chronic lymphocytic leukemia (CLL) is a B-cell-derived hematologic malignancy whose progression depends on active B-cell receptor (BCR) signaling. Despite the spectacular efficacy of Ibrutinib, an irreversible inhibitor of Bruton tyrosine kinase (BTK), resistance can develop in CLL patients, and alternative therapeutic strategies are therefore required. Cancer immunotherapy has revolutionized cancer care and may be an attractive approach in this context. We speculated that characterizing the immune responses of CLL patients may highlight putative immunotherapeutic targets. Here, we used high-dimensional spectral flow cytometry to compare the distribution and phenotype of non-B-cell immune populations in the circulating blood of CLL patients treated with Ibrutinib displaying a complete response or secondary progression. Although no drastic changes were observed in the composition of their immune subsets, the Ibrutinib-resistant group showed increased cycling of CD8+ T cells, leading to their overabundance at the expense of dendritic cells. In addition, the expression of 11 different surface checkpoints was similar regardless of response status. Together, this suggests that CLL relapse upon Ibrutinib treatment may not lead to major alterations in the peripheral immune response.


Subject(s)
Adenine , Immunophenotyping , Leukemia, Lymphocytic, Chronic, B-Cell , Piperidines , Humans , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Leukemia, Lymphocytic, Chronic, B-Cell/blood , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , Adenine/analogs & derivatives , Adenine/therapeutic use , Adenine/pharmacology , Piperidines/therapeutic use , Piperidines/pharmacology , Male , Female , Aged , Middle Aged , Aged, 80 and over , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/drug effects , Blood Cells/drug effects , Blood Cells/metabolism , Pyrimidines/therapeutic use , Pyrimidines/pharmacology , Drug Resistance, Neoplasm/drug effects
3.
Front Cell Infect Microbiol ; 14: 1413103, 2024.
Article in English | MEDLINE | ID: mdl-39113822

ABSTRACT

Background: Sepsis represents a severe manifestation of infection often accompanied by metabolic disorders and mitochondrial dysfunction. Notably, mitochondrial DNA copy number (mtDNA-CN) and the expression of specific mitochondrial genes have emerged as sensitive indicators of mitochondrial function. To investigate the utility of mitochondrial gene expression in peripheral blood cells for distinguishing severe infections and predicting associated outcomes, we conducted a prospective cohort study. Methods: We established a prospective cohort comprising 74 patients with non-sepsis pneumonia and 67 cases of sepsis induced by respiratory infections, aging from 2 to 6 years old. We documented corresponding clinical data and laboratory information and collected blood samples upon initial hospital admission. Peripheral blood cells were promptly isolated, and both total DNA and RNA were extracted. We utilized absolute quantification PCR to assess mtDNA-CN, as well as the expression levels of mt-CO1, mt-ND1, and mt-ATP6. Subsequently, we extended these comparisons to include survivors and non-survivors among patients with sepsis using univariate and multivariate analyses. Receiver operating characteristic (ROC) curves were constructed to assess the diagnostic potential. Results: The mtDNA-CN in peripheral blood cells was significantly lower in the sepsis group. Univariate analysis revealed a significant reduction in the expression of mt-CO1, mt-ND1, and mt-ATP6 in patients with sepsis. However, multivariate analysis did not support the use of mitochondrial function in peripheral blood cells for sepsis diagnosis. In the comparison between pediatric sepsis survivors and non-survivors, univariate analysis indicated a substantial reduction in the expression of mt-CO1, mt-ND1, and mt-ATP6 among non-survivors. Notably, total bilirubin (TB), mt-CO1, mt-ND1, and mt-ATP6 levels were identified as independent risk factors for sepsis-induced mortality. ROC curves were then established for these independent risk factors, revealing areas under the curve (AUCs) of 0.753 for TB (95% CI 0.596-0.910), 0.870 for mt-CO1 (95% CI 0.775-0.965), 0.987 for mt-ND1 (95% CI 0.964-1.000), and 0.877 for mt-ATP6 (95% CI 0.793-0.962). Conclusion: MtDNA-CN and mitochondrial gene expression are closely linked to the severity and clinical outcomes of infectious diseases. Severe infections lead to impaired mitochondrial function in peripheral blood cells. Notably, when compared to other laboratory parameters, the expression levels of mt-CO1, mt-ND1, and mt-ATP6 demonstrate promising potential for assessing the prognosis of pediatric sepsis.


Subject(s)
DNA, Mitochondrial , ROC Curve , Sepsis , Humans , Sepsis/blood , Sepsis/diagnosis , Sepsis/mortality , Child, Preschool , Female , Male , DNA, Mitochondrial/genetics , Prospective Studies , Prognosis , Child , Mitochondria/genetics , Mitochondria/metabolism , NADH Dehydrogenase/genetics , Mitochondrial Proton-Translocating ATPases/genetics , Blood Cells/metabolism , Genes, Mitochondrial , Gene Expression , Pneumonia/diagnosis , Pneumonia/blood , Predictive Value of Tests
4.
BMC Med Genomics ; 17(1): 212, 2024 Aug 14.
Article in English | MEDLINE | ID: mdl-39143590

ABSTRACT

Non-coding RNA expression has shown to have cell type-specificity. The regulatory characteristics of these molecules are impacted by changes in their expression levels. We performed next-generation sequencing and examined small RNA-seq data obtained from 6 different types of blood cells separated by fluorescence-activated cell sorting of severe COVID-19 patients and healthy control donors. In addition to examining the behavior of piRNA in the blood cells of severe SARS-CoV-2 infected patients, our aim was to present a distinct piRNA differential expression portrait for each separate cell type. We observed that depending on the type of cell, different sorted control cells (erythrocytes, monocytes, lymphocytes, eosinophils, basophils, and neutrophils) have altering piRNA expression patterns. After analyzing the expression of piRNAs in each set of sorted cells from patients with severe COVID-19, we observed 3 significantly elevated piRNAs - piR-33,123, piR-34,765, piR-43,768 and 9 downregulated piRNAs in erythrocytes. In lymphocytes, all 19 piRNAs were upregulated. Monocytes were presented with a larger amount of statistically significant piRNA, 5 upregulated (piR-49039 piR-31623, piR-37213, piR-44721, piR-44720) and 35 downregulated. It has been previously shown that piR-31,623 has been associated with respiratory syncytial virus infection, and taking in account the major role of piRNA in transposon silencing, we presume that the differential expression patterns which we observed could be a signal of indirect antiviral activity or a specific antiviral cell state. Additionally, in lymphocytes, all 19 piRNAs were upregulated.


Subject(s)
COVID-19 , Flow Cytometry , RNA, Small Interfering , SARS-CoV-2 , Humans , COVID-19/genetics , COVID-19/virology , RNA, Small Interfering/metabolism , RNA, Small Interfering/genetics , SARS-CoV-2/genetics , Male , Female , Middle Aged , Monocytes/metabolism , Adult , Blood Cells/metabolism , Piwi-Interacting RNA
5.
Zhonghua Yi Xue Za Zhi ; 104(33): 3087-3091, 2024 Aug 27.
Article in Chinese | MEDLINE | ID: mdl-39168838

ABSTRACT

The morphological examination of blood cells under manual microscopes is a classic method, but the obvious shortcomings limit the extensive development of peripheral blood cell morphological examination. By using the manual microscope method, it is difficult to ensure the effective implementation of reviewing rules on blood cell analysis, fails to meet the needs of clinical diagnosis and treatment activities, and exposes the risk of missing diagnosis and early detection of some blood system diseases. Artificial Intelligence-assisted peripheral blood cell morphological examination is an important development direction in blood cell morphological examination and diagnosis. This article primarily introduces the characteristics and current application status of artificial intelligence-assisted blood cell morphological examination, as well as the functional requirements and expectations for future technological advancements. Furthermore, it provides an outlook on potential clinical and laboratory application scenarios.With the rapid development of artificial intelligence in blood cell morphology testing, the performance of automated blood cell morphology analysis systems will be greatly improved, which can fully meet the needs of clinical diagnosis, treatment, and health management.


Subject(s)
Artificial Intelligence , Blood Cells , Humans , Blood Cells/cytology , Microscopy
6.
Clin Exp Med ; 24(1): 181, 2024 Aug 06.
Article in English | MEDLINE | ID: mdl-39105953

ABSTRACT

Traditional manual blood smear diagnosis methods are time-consuming and prone to errors, often relying heavily on the experience of clinical laboratory analysts for accuracy. As breakthroughs in key technologies such as neural networks and deep learning continue to drive digital transformation in the medical field, image recognition technology is increasingly being leveraged to enhance existing medical processes. In recent years, advancements in computer technology have led to improved efficiency in the identification of blood cells in blood smears through the use of image recognition technology. This paper provides a comprehensive summary of the methods and steps involved in utilizing image recognition algorithms for diagnosing diseases in blood smears, with a focus on malaria and leukemia. Furthermore, it offers a forward-looking research direction for the development of a comprehensive blood cell pathological detection system.


Subject(s)
Blood Cells , Image Processing, Computer-Assisted , Pathology, Clinical , Pathology, Clinical/methods , Pathology, Clinical/trends , Blood Cells/microbiology , Blood Cells/parasitology , Blood Cells/pathology , Malaria/diagnostic imaging , Leukemia/diagnostic imaging , Algorithms , Machine Learning , Blood Cell Count , Humans
7.
Zhonghua Xue Ye Xue Za Zhi ; 45(4): 330-338, 2024 Apr 14.
Article in Chinese | MEDLINE | ID: mdl-38951059

ABSTRACT

Blood cell morphological examination is a crucial method for the diagnosis of blood diseases, but traditional manual microscopy is characterized by low efficiency and susceptibility to subjective biases. The application of artificial intelligence (AI) technology has improved the efficiency and quality of blood cell examinations and facilitated the standardization of test results. Currently, a variety of AI devices are either in clinical use or under research, with diverse technical requirements and configurations. The Experimental Diagnostic Study Group of the Hematology Branch of the Chinese Medical Association has organized a panel of experts to formulate this consensus. The consensus covers term definitions, scope of application, technical requirements, clinical application, data management, and information security. It emphasizes the importance of specimen preparation, image acquisition, image segmentation algorithms, and cell feature extraction and classification, and sets forth basic requirements for the cell recognition spectrum. Moreover, it provides detailed explanations regarding the fine classification of pathological cells, requirements for cell training and testing, quality control standards, and assistance in issuing diagnostic reports by humans. Additionally, the consensus underscores the significance of data management and information security to ensure the safety of patient information and the accuracy of data.


Subject(s)
Artificial Intelligence , Blood Cells , Consensus , Humans , Blood Cells/cytology , China , Algorithms
8.
Phys Med Biol ; 69(17)2024 Aug 21.
Article in English | MEDLINE | ID: mdl-39084628

ABSTRACT

Purpose.To assess how inter-subject variations in brain vasculature among glioblastoma (GBM) patients affects the calculated dose received by circulating blood cells (CBC) during radiotherapy and its subsequent impact on CBC depletion.Methods.Ten GBM patients treated with either intensity-modulated radiation therapy (IMRT) or volumetric modulated arc therapy (VMAT) were selected. For each patient, 23 cerebrovascular models were developed based on 23 healthy subject MR-angiography data to simulate intra- and inter-subject blood vessel diversity. Based on the corresponding treatment plan of the patient, the dose to CBC was calculated for all the 230 scenarios. The impact of inter-subject variation on fraction of irradiated blood volume (VD>0 cGy) and lymphocyte kill rates as a function of the clinical target volume (CTV) size and treatment technique were analyzed.Results.The dose fluctuation to CBC was higher in IMRT plans compared to VMAT plans. The uncertainty in theVD>0 cGywas 18.3% for IMRT and 2.0% (CI95%) for VMAT and the dispersion of theD2%index was 6 cGy for IMRT and 1 cGy for VMAT (CI95%) for one single treatment fraction of 200 cGy. The uncertainty in killed CBC due to inter-subject diversity in brain blood vessel increased with increasing CTV size and wasσ= 11.2%.Conclusions. VMAT showed greater robustness against inter-subject variation in blood vessels compared to IMRT. We recommend considering the uncertainty in depleting CBC resulting from the use of less patient-specific and generic blood vessel phantoms to improve the radiation-induced lymphopenia assessments.


Subject(s)
Brain Neoplasms , Glioblastoma , Radiotherapy Dosage , Radiotherapy, Intensity-Modulated , Humans , Glioblastoma/radiotherapy , Uncertainty , Radiotherapy, Intensity-Modulated/adverse effects , Brain Neoplasms/radiotherapy , Radiotherapy Planning, Computer-Assisted/methods , Radiation Dosage , Blood Cells/radiation effects , Male
9.
Genome Res ; 34(7): 1089-1105, 2024 Aug 20.
Article in English | MEDLINE | ID: mdl-38951027

ABSTRACT

Knowledge of locations and activities of cis-regulatory elements (CREs) is needed to decipher basic mechanisms of gene regulation and to understand the impact of genetic variants on complex traits. Previous studies identified candidate CREs (cCREs) using epigenetic features in one species, making comparisons difficult between species. In contrast, we conducted an interspecies study defining epigenetic states and identifying cCREs in blood cell types to generate regulatory maps that are comparable between species, using integrative modeling of eight epigenetic features jointly in human and mouse in our Validated Systematic Integration (VISION) Project. The resulting catalogs of cCREs are useful resources for further studies of gene regulation in blood cells, indicated by high overlap with known functional elements and strong enrichment for human genetic variants associated with blood cell phenotypes. The contribution of each epigenetic state in cCREs to gene regulation, inferred from a multivariate regression, was used to estimate epigenetic state regulatory potential (esRP) scores for each cCRE in each cell type, which were used to categorize dynamic changes in cCREs. Groups of cCREs displaying similar patterns of regulatory activity in human and mouse cell types, obtained by joint clustering on esRP scores, harbor distinctive transcription factor binding motifs that are similar between species. An interspecies comparison of cCREs revealed both conserved and species-specific patterns of epigenetic evolution. Finally, we show that comparisons of the epigenetic landscape between species can reveal elements with similar roles in regulation, even in the absence of genomic sequence alignment.


Subject(s)
Epigenesis, Genetic , Epigenome , Species Specificity , Animals , Mice , Humans , Blood Cells/metabolism , Regulatory Sequences, Nucleic Acid , Gene Expression Regulation , Epigenomics/methods
10.
Methods Mol Biol ; 2796: 185-190, 2024.
Article in English | MEDLINE | ID: mdl-38856902

ABSTRACT

The potassium channels are one of the key regulators of cell membrane potential and permeability properties of blood cells. The changes in functioning of potassium channels control crucial cell processes such as proliferation, viability, migration, and invasion. The correct estimation of these processes is important for the characterization of physiological and pathophysiological cell states. Here, we present the experimental protocol for evaluation of the role of potassium ion channels in the proliferation, migration, and invasion of blood cells.


Subject(s)
Cell Movement , Cell Proliferation , Potassium Channels , Humans , Potassium Channels/metabolism , Blood Cells/metabolism , Blood Cells/cytology , Membrane Potentials
11.
Nature ; 631(8020): 281-283, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38926555
12.
J Nanobiotechnology ; 22(1): 363, 2024 Jun 24.
Article in English | MEDLINE | ID: mdl-38910248

ABSTRACT

Fluorescence nanoscopy, also known as super-resolution microscopy, has transcended the conventional resolution barriers and enabled visualization of biological samples at nanometric resolutions. A series of super-resolution techniques have been developed and applied to investigate the molecular distribution, organization, and interactions in blood cells, as well as the underlying mechanisms of blood-cell-associated diseases. In this review, we provide an overview of various fluorescence nanoscopy technologies, outlining their current development stage and the challenges they are facing in terms of functionality and practicality. We specifically explore how these innovations have propelled forward the analysis of thrombocytes (platelets), erythrocytes (red blood cells) and leukocytes (white blood cells), shedding light on the nanoscale arrangement of subcellular components and molecular interactions. We spotlight novel biomarkers uncovered by fluorescence nanoscopy for disease diagnosis, such as thrombocytopathies, malignancies, and infectious diseases. Furthermore, we discuss the technological hurdles and chart out prospective avenues for future research directions. This review aims to underscore the significant contributions of fluorescence nanoscopy to the field of blood cell analysis and disease diagnosis, poised to revolutionize our approach to exploring, understanding, and managing disease at the molecular level.


Subject(s)
Blood Cells , Microscopy, Fluorescence , Animals , Humans , Blood Cells/ultrastructure , Blood Platelets/metabolism , Erythrocytes , Hematology/methods , Leukocytes/metabolism , Microscopy, Fluorescence/methods , Nanotechnology/methods
13.
Free Radic Biol Med ; 222: 43-61, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38848784

ABSTRACT

There is a growing interest in using extracellular vesicles (EVs) for therapeutic applications. EVs are composed of cytoplasmic proteins and nucleic acids and an external lipid bilayer containing transmembrane proteins on their surfaces. EVs can alter the state of the target cells by interacting with the receptor ligand of the target cell or by being internalised by the target cell. Blood cells are the primary source of EVs, and 1 µL of plasma contains approximately 1.5 × 107 EVs. Owing to their easy acquisition and the avoidance of cell amplification in vitro, using blood cells as a source of therapeutic EVs has promising clinical application prospects. This review summarises the characteristics and biological functions of EVs derived from different blood cell types (platelets, erythrocytes, and leukocytes) and analyses the prospects and challenges of using them for clinical therapeutic applications. In summary, blood cell-derived EVs can regulate different cell types such as immune cells (macrophages, T cells, and dendritic cells), stem cells, and somatic cells, and play a role in intercellular communication, immune regulation, and cell proliferation. Overall, blood cell-derived EVs have the potential for use in vascular diseases, inflammatory diseases, degenerative diseases, and injuries. To promote the clinical translation of blood cell-derived EVs, researchers need to perform further studies on EVs in terms of scalable and reproducible isolation technology, quality control, safety, stability and storage, regulatory issues, cost-effectiveness, and long-term efficacy.


Subject(s)
Extracellular Vesicles , Humans , Extracellular Vesicles/metabolism , Cell Communication , Blood Cells/metabolism , Blood Cells/cytology , Animals , Erythrocytes/metabolism
14.
Article in English | MEDLINE | ID: mdl-38782370

ABSTRACT

Current therapies for acute radiation syndrome (ARS) involve bone marrow transplantation (BMT), leading to graft-versus-host disease (GvHD). To address this challenge, we have developed a novel donor-recipient chimeric cell (DRCC) therapy to increase survival and prevent GvHD following total body irradiation (TBI)-induced hematopoietic injury without the need for immunosuppression. In this study, 20 Lewis rats were exposed to 7 Gy TBI to induce ARS, and we assessed the efficacy of various cellular therapies following systemic intraosseous administration. Twenty Lewis rats were randomly divided into four experimental groups (n = 5/group): saline control, allogeneic bone marrow transplantation (alloBMT), DRCC, and alloBMT + DRCC. DRCC were created by polyethylene glycol-mediated fusion of bone marrow cells from 24 ACI (RT1a) and 24 Lewis (RT11) rat donors. Fusion feasibility was confirmed by flow cytometry and confocal microscopy. The impact of different therapies on post-irradiation peripheral blood cell recovery was evaluated through complete blood count, while GvHD signs were monitored clinically and histopathologically. The chimeric state of DRCC was confirmed. Post-alloBMT mortality was 60%, whereas DRCC and alloBMT + DRCC therapies achieved 100% survival. DRCC therapy also led to the highest white blood cell counts and minimal GvHD changes in kidney and skin samples, in contrast to alloBMT treatment. In this study, transplantation of DRCC promoted the recovery of peripheral blood cell populations after TBI without the development of GVHD. This study introduces a novel and promising DRCC-based bridging therapy for treating ARS and extending survival without GvHD.


Subject(s)
Acute Radiation Syndrome , Bone Marrow Transplantation , Disease Models, Animal , Graft vs Host Disease , Rats, Inbred Lew , Whole-Body Irradiation , Animals , Rats , Graft vs Host Disease/therapy , Graft vs Host Disease/immunology , Graft vs Host Disease/prevention & control , Bone Marrow Transplantation/methods , Acute Radiation Syndrome/therapy , Transplantation Chimera , Male , Transplantation, Homologous , Humans , Blood Cells
15.
Anat Histol Embryol ; 53(3): e13054, 2024 May.
Article in English | MEDLINE | ID: mdl-38735037

ABSTRACT

Identifying and analysing distinct blood cells is crucial for the diagnosis and treatment of diseases in the field of biomedicine. The present study was undertaken to study the cytomorphological and cytochemical characteristics of the blood cells of Zoar, a non-descript indigenous breed of chicken extensively reared under backyard poultry farming in Mizoram, India. For this study, 2 mL of blood samples were aseptically collected from the wings veins of 12 chickens and were processed for light microscopic study under standard protocols. The matured erythrocytes were elliptical, while the immature erythrocytes appeared oval. The heterophils were positive for SBB (SBB), Periodic Acid Schiff (PAS), acid phosphatase, alkaline phosphatase and Arylsulphatase while the eosinophils were positive for SBB, PAS, alkaline phosphatase, cytochrome oxidase and peroxidase. The basophils of were positive for toluidine blue while the thrombocytes were positive for PAS. These cytochemical and cytoenzymatic staining properties plays a very important role in diagnosis, differentiation, and classification of leukaemias.


Subject(s)
Chickens , Eosinophils , Erythrocytes , Animals , Chickens/anatomy & histology , India , Erythrocytes/cytology , Eosinophils/cytology , Blood Cells/cytology , Blood Platelets/cytology , Alkaline Phosphatase/blood , Basophils/cytology , Acid Phosphatase/blood , Electron Transport Complex IV/analysis
16.
J Vis Exp ; (205)2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38558008

ABSTRACT

Induced pluripotent stem cell (iPSC)-based models are excellent platforms to understand blood development, and iPSC-derived blood cells have translational utility as clinical testing reagents and transfusable cell therapeutics. The advent and expansion of multiomics analysis, including but not limited to single nucleus RNA sequencing (snRNAseq) and Assay for Transposase-Accessible Chromatin sequencing (snATACseq), offers the potential to revolutionize our understanding of cell development. This includes developmental biology using in vitro hematopoietic models. However, it can be technically challenging to isolate intact nuclei from cultured or primary cells. Different cell types often require tailored nuclear preparations depending on cellular rigidity and content. These technical difficulties can limit data quality and act as a barrier to investigators interested in pursuing multiomics studies. Specimen cryopreservation is often necessary due to limitations with cell collection and/or processing, and frozen samples can present additional technical challenges for intact nuclear isolation. In this manuscript, we provide a detailed method to isolate high-quality nuclei from iPSC-derived cells at different stages of in vitro hematopoietic development for use in single-nucleus multiomics workflows. We have focused the method development on the isolation of nuclei from iPSC-derived adherent stromal/endothelial cells and non-adherent hematopoietic progenitor cells, as these represent very different cell types with regard to structural and cellular identity. The described troubleshooting steps limited nuclear clumping and debris, allowing the recovery of nuclei in sufficient quantity and quality for downstream analyses. Similar methods may be adapted to isolate nuclei from other cryopreserved cell types.


Subject(s)
Cell Nucleus , Endothelial Cells , Cell Nucleus/metabolism , Cryopreservation/methods , Hematopoietic Stem Cells , Blood Cells
17.
Front Public Health ; 12: 1336674, 2024.
Article in English | MEDLINE | ID: mdl-38590804

ABSTRACT

Background: Hyperuricemia is a common metabolic disorder linked to various health conditions. Its prevalence varies among populations and genders, and high-altitude environments may contribute to its development. Understanding the connection between blood cell parameters and hyperuricemia in high-altitude areas can shed light on the underlying mechanisms. This study aimed to investigate the relationship between blood cell parameters and hyperuricemia in high-altitude areas, with a particular focus on gender differences. Methods: We consecutively enrolled all eligible Tibetan participants aged 18-60 who were undergoing routine medical examinations at the People's Hospital of Chaya County between January and December 2022. During this period, demographic and laboratory data were collected to investigate the risk factors associated with hyperuricemia. Results: Among the participants, 46.09% were diagnosed with hyperuricemia. In the male cohort, significant correlations were found between serum uric acid (SUA) levels and red blood cell (RBC) count, creatinine (Cr). Urea, alanine transaminase (ALT), and albumin (ALB). Notably, RBC exhibited the strongest association. Conversely, in the female cohort, elevated SUA levels were associated with factors such as white blood cell (WBC) count. Urea, ALT, and ALB, with WBC demonstrating the most significant association. Further analysis within the female group revealed a compelling relationship between SUA levels and specific white blood cell subtypes, particularly neutrophils (Neu). Conclusion: This study revealed gender-specific associations between SUA levels and blood cell parameters in high-altitude areas. In males, RBC count may play a role in hyperuricemia, while in females, WBC count appears to be a significant factor. These findings contribute to our understanding of metabolic dynamics in high-altitude regions but require further research for comprehensive mechanistic insights.


Subject(s)
Hyperuricemia , Humans , Male , Female , Hyperuricemia/epidemiology , Altitude , Uric Acid , Blood Cells , Urea
18.
BMC Med Imaging ; 24(1): 83, 2024 Apr 08.
Article in English | MEDLINE | ID: mdl-38589793

ABSTRACT

The research focuses on the segmentation and classification of leukocytes, a crucial task in medical image analysis for diagnosing various diseases. The leukocyte dataset comprises four classes of images such as monocytes, lymphocytes, eosinophils, and neutrophils. Leukocyte segmentation is achieved through image processing techniques, including background subtraction, noise removal, and contouring. To get isolated leukocytes, background mask creation, Erythrocytes mask creation, and Leukocytes mask creation are performed on the blood cell images. Isolated leukocytes are then subjected to data augmentation including brightness and contrast adjustment, flipping, and random shearing, to improve the generalizability of the CNN model. A deep Convolutional Neural Network (CNN) model is employed on augmented dataset for effective feature extraction and classification. The deep CNN model consists of four convolutional blocks having eleven convolutional layers, eight batch normalization layers, eight Rectified Linear Unit (ReLU) layers, and four dropout layers to capture increasingly complex patterns. For this research, a publicly available dataset from Kaggle consisting of a total of 12,444 images of four types of leukocytes was used to conduct the experiments. Results showcase the robustness of the proposed framework, achieving impressive performance metrics with an accuracy of 97.98% and precision of 97.97%. These outcomes affirm the efficacy of the devised segmentation and classification approach in accurately identifying and categorizing leukocytes. The combination of advanced CNN architecture and meticulous pre-processing steps establishes a foundation for future developments in the field of medical image analysis.


Subject(s)
Deep Learning , Humans , Data Curation , Leukocytes , Neural Networks, Computer , Blood Cells , Image Processing, Computer-Assisted/methods
19.
Front Immunol ; 15: 1329820, 2024.
Article in English | MEDLINE | ID: mdl-38590526

ABSTRACT

The immune system of Asian elephants (Elephas maximus) is poorly studied, compared to that of livestock, rodents or humans. The innate immune response has become a focus of interest in relation to Elephant endotheliotropic herpesviruses (EEHVs). EEHVs cause a fatal hemorrhagic disease (EEHV-HD) and are a significant threat to captive Asian elephant populations worldwide. Similar to other herpesvirus infections, nearly all animals become infected, but only some develop disease. As progression to EEHV-HD is often acute, a robust innate immune response is crucial to control EEHV infections. This is invariably true of the host in the first instance, but it can also potentially be modulated by intervention strategies. Here, two immunostimulant veterinary medicinal products, authorized for use in domestic species, were tested for their ability to induce innate anti-viral immune responses in Asian elephant blood cells. Sequence data were obtained for a range of previously unidentified Asian elephant immune genes, including C-X-C motif chemokine ligand 10 (CXCL10), interferon stimulated gene 15 (ISG15) and myxovirus GTPase 1 (Mx1), and were employed in the design of species-specific qPCR assays. These assays were subsequently used in analyses to determine fold changes in gene expression over a period of 24 hours. This study demonstrates that both immunostimulant medications are capable of inducing significant innate anti-viral immune responses which suggests that both could be beneficial in controlling EEHV infections in Asian elephants.


Subject(s)
Elephants , Herpesviridae Infections , Herpesviridae , Humans , Animals , Sheep , Elephants/genetics , DNA, Bacterial , Blood Cells , Immunity, Innate , Plasmids , Immunization , Adjuvants, Immunologic , Gene Expression
20.
Acta Neurobiol Exp (Wars) ; 84(1): 70-79, 2024 Mar 28.
Article in English | MEDLINE | ID: mdl-38587322

ABSTRACT

Hemorrhagic complications may be seen following reperfusion therapy with rtPA and/or thrombectomy after acute ischemic stroke (AIS). Neutrophils, lymphocytes, and platelets have important roles in the inflammatory and immune responses that develop in these patients. We investigated time­dependent changes in blood cells, NIHSS and mRS values according to type of reperfusion therapy in AIS patients who developed cerebral hemorrhage. In AIS patients who underwent rtPA and/or thrombectomy and developed cerebral hemorrhage within the first 24 hours after treatment, leukocyte, neutrophil, lymphocyte, platelet counts and their ratios were recorded on admission, 1st, 3rd, and 7th days. NIHSS values on admission, 3rd days and mRS values on admission, discharge, and the 3rd month were recorded. These values were compared according to the type of reperfusion therapy. Out of 436 AIS patients, rtPA was applied in 50.5%, thrombectomy in 28.2%, and rtPA+thrombectomy in 21.3%. Hemorrhage developed in 25.5% of the patients. Patients treated with thrombectomy had a greater rate of cerebral hemorrhage. Pre­stroke mRS values were lower in all therapy types than mRS scores at discharge and the 3rd month. The NIHSS scores did not differ significantly in 3 days. Depending on the type of reperfusion treatment, there are a few time­dependent significant changes observed in the blood cell counts and ratios. In conclusion, there is a relation between the type of reperfusion therapy and the time­dependent changes in blood cells and ratios as well as mRS scores among AIS patients who have undergone rtPA and/or thrombectomy and developed cerebral hemorrhage.


Subject(s)
Brain Ischemia , Ischemic Stroke , Stroke , Humans , Ischemic Stroke/complications , Ischemic Stroke/therapy , Treatment Outcome , Stroke/complications , Stroke/therapy , Cerebral Hemorrhage/complications , Blood Cells , Reperfusion/adverse effects , Brain Ischemia/complications
SELECTION OF CITATIONS
SEARCH DETAIL