Healthy Blood Samples Research Articles

Identification of Some Fungi Causing Skin Infection and its Relationship with Some Blood Test of Tinea Patients in Tikrit City

Dermatophytes infection is a common problem worldwide and frequent in Iraq. Several reports and articles were published on prevalence, distribution, causes and treatment of dermatophytosis. The current study included the collection of 100 clinical samples (70 skin peels, 20 hair samples and 10 nail samples), as well as 50 blood samples from patients with dermatophytes and skin consultants in Tikrit City and 20 healthy blood samples for the period from November 2023 to March 2024. The results showed that the most common dermatophytes isolated was Microsporum canis and T.rubrum 10 (20 %) isolates from 50 total , followed by M.gypseum, 6(12%) and 5(10%) T.mentagrophytes and T.verrucosum and 4(8 %) isolates E. floccosum and T.erinacei and finally appeared 3(6 %) isolate T.schoenleinii and T.concentricum. The results of the statistical analysis showed non significant difference P <0.05 in the total number of white blood cells and monocyte (5.15 ± 2.636, 0.89±0.51) in patients with dermatophytes compared with the group Control mean± SD (5.73±1.821, ± 0.69 ± 1.591) respectively. Lymphocytes significant decreased (1.82 ± 1.301) in patients with dermatophytes compared to control group (4.60 ± 1.35) P≤0.01. Furthermore granlocytes increased (2.30±1.591) in patients with dermatophytes compared to control group (0.33± 0.403) P<0.01. The study concluded that perhaps the reduction of some of the body's WBCs parameters has an important role in the incidence of fungal infection in people infected with them when compared to healthy people.

Affinity-based 3D-printed microfluidic chip for clinical sepsis detection with CD69, CD64, and CD25

Sepsis is a life-threatening immune response to infection in the body, eventually resulting in fatal organ failure. Current methods utilize blood cultures and quick-Sequential-Organ-Failure-Assessment (qSOFA), but there is a need for more accurate and time-sensitive diagnostic methods to improve survival rates. We present a 3D-printed microfluidic chip that bioconjugates antibodies CD69, CD64, and CD25 to channel surfaces to capture sepsis cells in blood samples and validate it with clinical samples (n = 125 septic, n = 10 healthy). Other variables were taken such as healthy volunteer blood samples and patient demographics to validate and confirm our device’s diagnostic ability. Statistical differences were found between healthy volunteer and sepsis patient antigen cell counts (CD69 p-value < 0.001, CD64 p-value < 0.004, CD25 p-value < 0.0009), and were confirmed using principal component analysis. Demographics such as length of stay, age, culture results, and need for surgery also factored into sepsis detection on a smaller scale than the antigen cell counts. The receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) of 0.989, 0.988, and 0.992 for CD69, CD64, and CD25, respectively, and a combined biomarker panel of 0.997. Overall, the device performed within a shorter time frame of 4 h compared to standard blood culture tests and was validated for use in detecting sepsis in patients.

Fabrication of a cell irradiation technique by alpha-particles using allyl diglycol carbonate (ADC) detector and micro-capillary tubes.

To study the impact of micro-alpha irradiation collimator with a specific design to irradiate specific normal or up-normal cells using capillary tubes and nuclear track detector type CR-39. The in vitro experimental study was conducted at radiobiology Lab, of the Physics department, Salahaddin University, Erbil, Iraq from February to April 2022. Several alpha irradiation collimators were calibrated using allyl diglycol carbonate to fabricate a suitable blood cell irradiation technique. Time of irradiation and alpha particle energy were calibrated. Healthy blood samples of Albino rats and cancer blood samples were used to assess the applicability of the fabricated cell irradiation technique. The Rats were divided into intervention and control groups. Data was analysed using SPSS software version 28. Of the 15 healthy, male Albino rats having a mean weight of 230±12g each, there were 12(80%) in the intervention group and 3(20%) in the control group. Microcapillary tubes with suitable diameters had high stability for deposition of a sufficient density of alpha particles on the surface of allyl diglycol carbonate and blood samples. The optimum time of irradiation that had a significant (p<0.05) effect was 20 sec corresponding to alpha energy 4.5MeV. Low irradiation time had significant impact of alpha particles on the average percentage of lymphocyte and neutrophil cells.

Stromal-Like Cells Are Found in Peripheral Blood of Patients With Inflammatory Bowel Disease and Correlate With Immune Activation State.

Recent studies have identified a critical role of stromal-immune cell interactions in immunity and immune tolerance. Transcriptomic profiling has implicated stromal cells in immune-mediated disorders including the 2 common forms of inflammatory bowel disease (IBD), Crohn's disease (CD), and ulcerative colitis (UC). Stromal-immune interactions may edify inflammatory state and the development of IBD-related complications such as fibrosis, yet the lack of protein markers has hampered studying stromal-immune perturbation. In this study, we designed a 40-color spectral flow cytometry assay to characterize hematopoietic and nonhematopoietic cells in intestinal biopsies and matched blood samples from patients with CD or UC. We identified circulating stromal-like cells that are significantly more abundant in IBD blood samples than in healthy controls. Those cells expressed podoplanin (PDPN), a commonly used marker for fibroblasts, and they were associated with activated and memory T and B cells and altered natural killer cell, monocyte, and macrophage populations. PDPN + cells in the blood correlated with PDPN + cells in the colon. Principal component analysis distinctly separated healthy blood samples from IBD blood samples, with stromal-like cells and B-cell subtypes dominating the IBD signature; Pearson correlation detected an association between PDPN + stromal-like cells and B-cell populations in IBD blood and gut biopsies. These observations suggest that PDPN + cells in the blood may serve as a biomarker of IBD. Understanding the relationship between stromal cells and immune cells in the intestine and the blood may provide a window into disease pathogenesis and insight into therapeutic targets for IBD.

Возрастная динамика субпопуляций Т-лимфоцитов у пациентов с синдромом делеции 22q11.2 (синдромом Ди Джорджи)

Patients with the 22q11.2 deletion syndrome (DiGeorge syndrome, DDS) suffer from T-cell lymphopenia, changes in function and subpopulation of T-lymphocytes. The bibliographical data conflicts on the characteristics of T-lymphocytes in patients with DDS and their changes during physiological maturation. The purpose of this research was to follow-up patients with emphasis on the dynamics of changes in T-lymphocyte subpopulations with age. Methods used: 117 patients observed during 2013-2023 who were administered at the Allergology and Immunology Department with the G.N. Speransky City Children’s Hospital No. 9 (Moscow, Russia) with a diagnosis of DDS and who had undergone blood sampling to assess T-lymphocyte subpopulations using flow cytometry. All patients (0 to 18 y/o) were divided into 4 age groups: 0 to 2 y/o, 2 to 5 y/o, 5 to 10 y/o and 10 to 18 y/o. In parallel, the blood samples of 185 apparently healthy children of the corresponding age were examined. Results: a decrease in CD3 T-lymphocytes, CD4 T-helper cells, CD8 T-cytotoxic lymphocytes, CD4 early thymic emigrants and CD4 naive T-lymphocytes was found in all age groups (p&lt;0.05). CD8+ naive T-lymphocytes and CD8+ early thymic emigrants were reduced in all age groups, but in the 2 to 5 y/o age group their values were close to normal (p=0.072 and p=0.220, respectively). CD4+ central memory cells were elevated in all age groups (p&lt;0.001), while CD8+ central memory cells, CD4+ and CD8+ effector memory cells differed in that they had normal values in the 2 to 5 y/o age group (p=0.229, p=0.457 and p=0.140, respectively). CD4+ TEMRA were significantly increased in the 0 to 2 y/o age group (p=0.002), and in older age groups they tended to normal values. CD8+ TEMRA did not differ from the control group and did not have age-related dynamics. The T-helper subpopulations were characterized by a significant increase in the percentage of T-helper type 1, T-helper 17, T-helper 17.1 and a decrease in T-helper type 2 compared to the population of healthy blood sample donors (p=0.001), except for patients from the 2 to 5 y/o age group. T-regulatory cells in absolute amount were reduced in all age groups (p&lt;0.001), while their relative number corresponded to the norm in the 0 to 2 y/o age group (p=0.811) and decreased slightly in patients in older age groups, especially in the 2 to 5 y/o age group (p=0.030). Conclusion: the pathology of thymus development in patients with DDS leads to impaired maturation of T-lymphocytes, leading to an increase in the number of mature forms of T-lymphocytes, shifts towards the development of T-helper 1 and T-helper 17, and a decrease in T-regulatory cells. Disturbances in T-lymphocytes can cause changes (dysregulation) in subpopulations of B-cells. All these processes may underlie the progression of autoimmune and infectious complications in such patients that are increasing with aging.

Transcriptomic signature of CD4-expressing T-cell abundance developed in healthy peripheral blood predicts strong anti-retroviral therapeutic response in HIV-1: A retrospective and proof-of-concept study

CD4-expressing T-cells (CD4Ts) play a crucial role in maintaining the normal functioning of the mammalian immune system and overall systemic health. Diseased individuals, such as those infected with the human immunodeficiency virus type I (HIV-1), experience progressive and eventual depletion of CD4T leading to uncurable conditions and ultimate death if left untreated. Although much is known about the role of CD4T-mediated immunity, the understanding of CD4T-related transcriptomic patterns remains incomplete. This proof-of-concept study aims to identify a transcriptome-wide gene signature for CD4T abundance by Least Absolute Shrinkage and Selection Operator (LASSO) regression modeling in 340 healthy peripheral blood samples. The optimized LASSO model demonstrated computational robustness (tenfold average Pearson&amp;rsquo;s r = 0.89) and biological relevance evidenced by four significant Gene Ontology terms (all odds ratio [OR] &amp;ge; 4.5 and false discovery rate &amp;le;0.05). Subsequently, in an independent cohort with 24 HIV-1-infected men who received anti-retroviral therapies, there is a significant, positive association between the gene signature and a strong anti-retroviral response before (OR = 13.6, P &lt; 0.05) and after adjusting for subject age, sex, and race (OR = 14.4, P &lt; 0.05). Taken together, the gene expression pattern associated with CD4T abundance is predictable, generalizable, and biologically relevant, shedding new light on the importance of CD4T abundance.

Aggregation-Augmented Magnetism of Lanthanide-Doped Nanoparticles and Enabling Magnetic Levitation-Based Exosome Sensing.

Due to the presence of unpaired electron orbitals in most lanthanide ions, lanthanide-doped nanoparticles (LnNPs) exhibit paramagnetism. However, as to biosensing applications, the magnetism of LnNPs is so weak that can hardly be employed in target separation. Herein, it is discovered that the magnetism of the LnNPs is highly associated with their concentration in a confined space, enabling aggregation-augmented magnetism to make them susceptive to a conventional magnet. Accordingly, a magnetic levitation (Maglev) sensing system is designed, in which the target exosomes can specifically introduce paramagnetic LnNPs to the microbeads' surface, allowing aggregation-augmented magnetism and further leverage the microbeads' levitation height in the Maglev device to indicate the target exosomes' content. It is demonstrated that this Maglev system can precisely distinguish healthy people's blood samples from those of breast cancer patients. This is the first work to report that LnNPs hold great promise in magnetic separation-based biological sample sorting, and the LnNP-permitted Maglev sensing system is proven to be promising for establishing a new generation of biosensing devices.

Characterization of T-cell receptor repertoire and correlation with tumor mutational landscape in lung cancer.

2577 Background: T-cell receptor (TCR) repertoire represents an overall immune condition which is closely related to pathogens- or tumor-associated T cell responses. Increasing evidence suggests that TCR repertoire is expected to undergo cancer-specific changes during tumorigenesis, supporting that TCR characteristics can serve as novel markers to indicate early cancer progression. Here, we performed a systematic T cell repertoire analysis in lung cancer. We identified cancer-enriched TCR signatures and constructed a comprehensive lung cancer database integrating TCR diversity, tumor mutational profiles, immune markers expression and HLA genotypes. Methods: A total of 988 tissue samples and 3,360 blood samples were obtained from patients with lung cancer, in addition to 2,699 blood samples from healthy individuals. Multiplex-PCR-based sequencing of the CDR3 regions of TCR-β chains was applied to the tissue and blood samples. TCR repertoires were analyzed using MiXCR and VDJtools. Whole exome sequencing was performed on partial tumor tissues to profile the mutational landscape and HLA genotypes. An in-house pipeline, considering the frequency and distribution of CDR3, was utilized to identify cancer-enriched CDR3 sequences. The PD-L1 expression was evaluated and quantified using CPS. Furthermore, an enrich score was developed to measure the content of cancer-related TCRs by aligning against the built cancer-enriched CDR3 dataset. Spearman's rank correlation was used to assess relationships between variables. Results: Lung cancer exhibited significantly lower Shannon index, Simpson index and evenness index in TCR repertoire (p&lt;0.001), both in tissue and blood samples, when compared with healthy blood samples. Approximate 3% T-cell clonotypes within lung tissues were detected in blood TCR repertoires. A correlation was observed between the decreased TCR diversity and a higher tumor mutational burden (TMB)(r = -0.15, p &lt; 0.01), as well as a higher variant allele frequency (VAF) (r = -0.19, p &lt; 0.01). In this study, 3,652 and 3,840 CDR3 sequences were identified to be enriched in lung cancer tissue and blood respectively. A significant difference in cancer-enriched score was observed between cancer and healthy TCR repertoires (p &lt; 0.001). The cancer-enriched score showed a significant positive correlation with TMB, particularly in TP53- mutant tumors (r = 0.27, p &lt; 0.001). A higher CPS (PD-L1) was associated with a higher cancer-enriched score (r = 0.18 ,p &lt; 0.01). Conclusions: Cancer-related T-cell clonotypes in the tumor microenvironment and peripheral blood informs the changes in anti-tumor responses, supporting the potential application of TCR signatures in early cancer detection. This study identified the lung cancer-related TCR signatures and provided a comprehensive lung cancer TCR database, revealing the relationship between TCR characteristics and mutational profiles. Clinical trial information: ChiCTR2200055761.

Evaluation of Fibrin Clot Interaction with Dental Implant after Different Surface Treatments: An In Vitro Study.

The current study was carried out to assess the interaction between fibrin clots and dental implants following various surface treatments. In this investigation, 45 dental implants with dimensions of 16 mm in length and 5 mm in diameter were utilized. They were divided up into three groups, each consisting of fifteen samples. Group I: Control; Group II: Ultraviolet (UV) light treated; and group III: Sandblasted and acid-etching (SLA) treated. Healthy volunteers' venous blood samples were drawn into vacutainer tubes without the use of anticoagulants. The samples were centrifuged for 3 minutes at 2700 rpm in a table centrifuge. The entire implant was submerged in room-temperature liquid fibrinogen for 60 minutes. Then, scanning electronic microscopy (SEM) was used to examine each sample. The inter- and intragroup assessments were obtained using the Mann-Whitney U test and the Kruskal-Wallis test; p-values less than 0.05 were regarded as statistically significant. The maximum adhesion of fibrin clot was found in SLA treated group (2.42 ± 0.10) followed by the UV light-treated group (2.18 ± 0.08) and control group (1.20 ± 0.02). There was a statistically significant difference found between the three surface-treated groups (p < 0.001). All surface-treatment methods exhibit adhesion between the implant surface and the fibrin clot. However, the highest adherence of fibrin clot was found in SLA treated group compared to the UV light-treated and control group. The physical and chemical characteristics of an implant's surface have a significant impact on the way blood clots organize. At the interface between the implant and the bone, blood clot production can initiate and facilitate the healing process. How to cite this article: Jalaluddin M, Ramanna PK, Swain M, et al. Evaluation of Fibrin Clot Interaction with Dental Implant after Different Surface Treatments: An In Vitro Study. J Contemp Dent Pract 2024;25(3):276-279.

Determining the Reference Range of Amino Acids in Healthy Neonatal Blood Samples in Northeast Iran Using LC-MS/MS

Determining the Reference Range of Amino Acids in Healthy Neonatal Blood Samples in Northeast Iran Using LC-MS/MS

Comparison of results obtained using clot-fibrinolysis waveform analysis and global fibrinolysis capacity assay with rotational thromboelastography

Global fibrinolysis assays detect the fibrinolysis time of clot dissolution using tissue-type plasminogen activator (tPA). Two such assays, clot-fibrinolysis waveform analysis (CFWA) and global fibrinolysis capacity (GFC) assay, were recently developed. These were compared with rotational thromboelastography (ROTEM). Healthy donor blood samples were divided into four groups based on tPA-spiked concentrations: 0, 100, 500, and 1000 ng/mL. CFWA and GFC fibrinolysis times, including 4.1 µg/mL and 100 ng/mL tPA in the assays, were determined, denoted as CFWA-Lys and GFC-Lys, respectively. Statistical differences were recognized between tPA concentrations of 0 and 500/1000 ng/mL for CFWA-Lys, and 0 and 100/500/1000 ng/mL for GFC-Lys. The correlation coefficients with lysis onset time (LOT) of extrinsic pathway evaluation and intrinsic pathway evaluation in ROTEM were statistically significant at 0.610 and 0.590 for CFWA-Lys, and 0.939 and 0.928 for GFC-Lys, respectively (p-values < 0.0001 for all correlations). Both assays showed significant correlations with ROTEM; however, the GFC assay proved to have better agreement with ROTEM compared with the CFWA assay. These assays have the potential to reflect a hyperfibrinolysis status with high tPA concentrations.

Abstract 2304: Detection of early-disseminated cancer cells with deep learning-enabled holographic imaging

Abstract Recent research increasingly validates the early dissemination of tumor cells in peripheral blood during the nascent stages of cancer, often preceding clinical identification of the primary tumor. This emerging evidence emphasizes the potential of Circulating Tumor Cells (CTCs) as early indicators for cancer. Despite their significance, CTCs are exceptionally rare compared to normal blood cells, which presents a considerable challenge in detection, particularly during early stages of cancer. Traditional detection methods, such as those based on nucleated cell enrichment, antibody labeling and fluorescence imaging, encounter notable limitations when applied in the context of early detection. These limitations arise from the inherent heterogeneity of CTCs, and the multistep process involved in labeling and manual enumeration, compromising cell viability and resulting in cell loss. This loss becomes particularly problematic in the context of early cancer detection scenarios where the tumor burden is extremely low, and any cell loss can significantly impact assay sensitivity. To overcome these challenges, we propose an innovative CTC detection approach employing a deep learning framework combined with holographic imaging. This technique utilizes coherent light to generate holograms from cell samples, capturing detailed 3D morphological and optical properties of individual cells. We developed a custom convolutional neural network designed to facilitate high-throughput cell sorting in real-time. Trained on healthy blood samples and diverse cancer cell lines, the dataset consists of more than 50 million cellular holograms enhanced by proprietary Gaussian annotation and image augmentation strategies. Moreover, by combining the morphological signatures from holography with antigen signatures (e.g. HER2/ER/PR) in real-time, our model demonstrates the ability to detect less than 1 false positive per 1 million nucleated cells, highlighting its potential as a robust tool for targeted early-stage breast cancer detection. Our technology represents highly sensitive alternative to traditional screening methods, with the promise of enhancing patient outcomes. Citation Format: Nicholas Heller, Kevin Mallery, Nathaniel Bristow, Yash Travadi, Eng Hock Lee, Jiarong Hong. Detection of early-disseminated cancer cells with deep learning-enabled holographic imaging [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2304.

Abstract 7657: Detection of KMT2A-PTDs and KMT2A fusions using next generation sequencing

Abstract Introduction: KMT2A (MLL) rearrangements (fusions) is a therapeutic biomarker for Menin inhibitors. KMT2A-PTDs (partial tandem duplications) are considered a prognostic biomarker in myeloid malignancies. KMT2A fusions and PTDs are traditionally detected by RT-qPCR (quantitative real-time PCR). This study investigates KMT2A PTDs levels in healthy donors and myeloid malignancy samples to establish a threshold to report samples with high PTDs, using next generation sequencing (NGS) with OncomineTM Myeloid Assay GX v2. We also report KMT2A fusions in myeloid malignancies. Methods: We sequenced 8483 research samples with known myeloid malignancies at Sonora Quest LaboratoriesTM and at Thermo Fisher ScientificTM. We acquired 20 healthy donor whole blood samples (total 127 replicates) from StanfordTM Blood Center and Discovery Life SciencesTM and sequenced them at 3 different sites of Thermo Fisher ScientificTM. Samples were sequenced with the Ion TorrentTM GenexusTM 6.6 or Ion GeneStudioTM S5 System. They were profiled for 6 different KMT2A-PTD variants and 199 KMT2A fusion isoforms. Results: The mean read length of this data set is 90 - 120 bp and the mean mapped fusion reads is 20,000 - 30,000. KMT2A-PTDs were detected in both healthy donors and myeloid samples. Healthy donor PTD read counts were consistently &amp;lt;2000 and averaged 1/3 of myeloid sample PTD read counts. About 33% of myeloid samples had higher PTD read counts than any healthy donor sample. BLAT (BLAST-Like Alignment Tool) analysis confirmed specific exon matching on the KMT2A gene in both cohorts. Among the 8483 myeloid samples, 162 samples contained a total of 5 unique KMT2A PTDs, and 105 samples contained a total of 30 unique KMT2A fusion isoforms with KMT2A-MLLT1 and KMT2A-MLLT3 being the most prevalent KMT2A fusion gene pairs. Conclusions: We characterize the KMT2A fusions present in myeloid malignant samples. We also describe the abundance of KMT2A PTDs in both healthy donor and myeloid samples, with myeloid cases showing significantly higher PTD read counts. KMT2A PTD read count &amp;gt;2000 is present only in malignant samples but not in healthy donors. This intriguing finding opens opportunities for prospective studies to monitor individuals with elevated PTD levels for myeloid malignancy development and retrospective studies to explore whether healthy donors identified with this alteration years ago after blood donation were subsequently recorded in the national health system with myeloid malignancies. (For research use only. Not for use in diagnostic procedures. © 2023 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Stanford is a trademark of the Board of Trustees of the Leland Stanford Junior University. Discovery Life Sciences is a trademark of Discovery Life Sciences. Sonora Quest Laboratories is a trademark of Sonora Quest Laboratories.) Citation Format: Jiajie Huang, Haigang Gu, Janet Orton, Marina Sedova, Amir Marcovitz, Jennifer Burke, Sarah Brozio, Paul Williams, Scott Myrand, Nate Olowo, Adam Broomer, Brendan Deal, Collyn Seeger, Seth Sadis, Sophie Rozenzhak, Fiona Hyland, Guang Liu. Detection of KMT2A-PTDs and KMT2A fusions using next generation sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7657.

Upregulation of lnc-FOXD2-AS1, CDC45, and CDK1 in patients with primary non-M3 AML is associated with a worse prognosis

Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy with an unfavorable outcome. The present research aimed to identify novel biological targets for AML diagnosis and treatment. In this study, we performed an in-silico method to identify antisense RNAs (AS-RNAs) and their related co-expression genes. GSE68172 was selected from the AML database of the Gene Expression Omnibus and compared using the GEO2R tool to find DEGs. Antisense RNAs were selected from all the genes that had significant expression and a survival plot was drawn for them in the GEPIA database, FOXD2-AS1 was chosen for further investigation based on predetermined criteria (logFC ≥|1| and P < 0.05) and its noteworthy association between elevated expression level and a marked reduction in the overall survival (OS) in patients diagnosed with AML. The GEPIA database was utilized to investigate FOXD2-AS1-related co-expression and similar genes. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and gene ontology (GO) function analysis of the mentioned gene lists were performed using the DAVID database. The protein–protein interaction (PPI) network was then constructed using the STRING database. Hub genes were screened using Cytoscape software. Pearson correlation analysis was conducted using the GEPIA database to explore the relationship between FOXD2-AS1 and the hub genes. The transcription of the selected coding and non-coding genes, including FOXD2-AS1, CDC45, CDC20, CDK1, and CCNB1, was validated in 150 samples, including 100 primary AML non-M3 blood samples and 50 granulocyte colony stimulating factor (G-CSF)-mobilized healthy donors, using quantitative Real-Time PCR (qRT-PCR). qRT-PCR results displayed significant upregulation of lnc-FOXD2-AS1, CDC45, and CDK1 in primary AML non-M3 blood samples compared to healthy blood samples (P = 0.0032, P = 0.0078, and P = 0.0117, respectively). The expression levels of CDC20 and CCNB1 were not statistically different between the two sets of samples (P = 0.8315 and P = 0.2788, respectively). We identified that AML patients with upregulation of FOXD2-AS1, CDK1, and CDC45 had shorter overall survival (OS) and Relapse-free survival (RFS) compared those with low expression of FOXD2-AS1, CDK1, and CDC45. Furthermore, the receiver operating characteristic (ROC) curve showed the potential biomarkers of lnc -FOXD2-AS1, CDC45, and CDK1 in primary AML non-M3 blood samples. This research proposed that the dysregulation of lnc-FOXD2-AS1, CDC45, and CDK1 can contribute to both disease state and diagnosis as well as treatment. The present study proposes the future evolution of the functional role of lnc-FOXD2-AS1, CDC45, and CDK1 in AML development.

Orthostatic Hypotension Promotes the Progression From Mild Cognitive Impairment to Dementia in Type 2 Diabetes Mellitus.

In type 2 diabetes mellitus (T2DM), orthostatic hypotension (OH) is associated with cognition, but the mechanisms governing the link between OH and cognition are still unclear. We sought to analyze Alzheimer's disease (AD) biomarkers and the part of complement proteins in modulating the association of OH with cognitive impairment and examine whether OH could accelerate the clinical progression of mild cognitive impairment (MCI) to dementia in T2DM. We recruited patients with T2DM with MCI and collected general healthy information and blood samples. Complement proteins of astrocyte-derived exosomes were isolated and AD biomarkers of neuronal cell-derived exosomes isolated were quantified by enzyme-linked immunosorbent assay. Cognitive assessments were performed at patient enrollment and follow-up. Mediation analysis showed that the influence of OH on cognition in T2DM was partly mediated by baseline AD biomarkers and complement proteins. Cox proportional-hazards regression proved the OH group had a higher risk of developing dementia compared to the T2DM without OH group. In T2DM with MCI patients, AD biomarkers and complement proteins mediate the effects of OH on cognitive impairment and OH may be a risk factor of progression from MCI to dementia in T2DM.

Study of Machine Learning Algorithms on Early Detection of Leukemia

Leukemia is a type of cancer that impacts the blood cells and the bone marrow it can be classified into acute and chronic. Early detection is crucial for reducing mortality rates. Acute leukemia progresses rapidly, causing bleeding issues, infections, and anemia due to premature of white blood cells accumulating blood vessels in bone marrow. Chronic leukemia advances slower, leading to an aggregation of abnormal cells. Detecting leukemia more effectively and accurately, automated and machine learning algorithms are being developed. Training algorithms on extensive blood smear images datasets allow these methods to differentiate normal cells from abnormal ones. With faster and more standardized detection of leukemia, medical pathologist can make more informed decisions. While machine learning algorithms may improve detection, skilled healthcare professionals remain essential for interpreting results and providing optimal patient care. The proposed model give a demonstration of a convolutional neural network (CNN) and TensorFlow framework, this method predicts leukemia cells from healthy blood samples this technique has gained popularity as a valuable tool for diagnosing leukemia as well as treating its accuracy of 92.62%. The second approach is that to classify large dataset images of the malignant cells from the normal cells, we employ a VGG19, a ResNet50, and a ResNet101 neural network, as well as batch normalization of the images achieved better accuracy and F1-score.

Investigation Roles of Erythropoietin, Testosterone, and Thyroid Hormones in Patients with Chronic Liver Disease

Chronic liver disease (CLD) is characterized as a steady decline in liver functions that lasts longer than six months, including the generation of clotting factors and other proteins, detoxification of toxic metabolic products, and bile excretion. CLD is a continual process of inflammation, damage, and regeneration of the liver parenchyma that results in fibrosis and cirrhosis. The study aims to determine the predictive role of erythropoietin, testosterone, and thyroid function markers in the pathogenesis of liver dysfunction in Iraqi patients. The current research investigation was conducted out in Baquba Teaching Hospital / Diyala governorate from November 2022 to January 2023. 50 blood samples were taken from patients who came to the Baquba Teaching Hospital and those with chronic liver disease for inspection and diagnosis by the consultant doctor in the advisory units/Baquba Teaching Hospital. 30 healthy people's blood samples were taken as a control group. The serum levels of erythropoietin, testosterone, TSH, FT3, and FT4 indicators in the samples were determined using the Roche Cobas e411. The current study's findings revealed that 61-70 and &gt;70 years scored highest age groups (28% and 26%) than ≤40 years that scored least age groups (6%) with significant differences (p&lt;0.05). The levels of erythropoietin and TSH were higher in patients than healthy. In contrast, the levels of testosterone, FT3, and FT4 were low in patients than healthy with significant differences (p&lt;0.05). According to Pearson correlations, erythropoietin is a substantial positive association with FT4 (r= 0.293* Sig.=0.039). Depending on receiver operating characteristic (ROC) curve, the present study showed the Erythropoietin, Testosterone, TSH, FT3, and FT4 markers scored the highest sensitivity (86%, 90%, 94%, 96%, and 100%) and specificity (90%, 90%, 94%, 100%, and 72%) respectively, in screening patients with Chronic liver disease (CLD). We came to the conclusion that illness severity increased with age. Erythropoietin, testosterone, and thyroid function are good prognostic markers in screening chronic liver disease that is associated with gonadal and thyroid disorders.

Proximal tubule cells in blood and urine as potential biomarkers for kidney disease biopsy

Early diagnosis and treatment are crucial for managing kidney disease, yet there remains a need to further explore pathological mechanisms and develop minimally invasive diagnostic methods. In this study, we employed single-cell RNA sequencing (scRNA-seq) to assess the cellular heterogeneity of kidney diseases. We analyzed gene expression profiles from renal tissue, peripheral blood mononuclear cells (PBMCs), and urine of four patients with nephritis. Our findings identified 12 distinct cell subsets in renal tissues and leukocytes. These subsets encompassed fibroblast cells, mesangial cells, epithelial cells, proximal tubule cells (PTCs), and six immune cell types: CD8+ T cells, macrophages, natural killer cells, dendritic cells, B cells, and neutrophils. Interestingly, PTCs were present in both PBMCs and urine samples but absent in healthy blood samples. Furthermore, several populations of fibroblast cells, mesangial cells, and PTCs exhibited pro-inflammatory or pro-apoptotic behaviors. Our gene expression analysis highlighted the critical role of inflammatory PTCs and fibroblasts in nephritis development and progression. These cells showed high expression of pro-inflammatory genes, which could have chemotactic and activating effect on neutrophils. This was substantiated by the widespread in these cells. Notably, the gene expression profiles of inflammatory PTCs in PBMCs, urine, and kidney tissues had high similarity. This suggests that PTCs in urine and PBMCs hold significant potential as alternative markers to invasive kidney biopsies.

High-throughput sequencing reveals the change of TCR α chain CDR3 with Takayasu arteritis.

Takayasu arteritis (TAK) is an inflammatory disease of blood vessels, and its pathogenesis is not clear at present. In this study, we explored the immunological characteristics of T cell receptor (TCR) α-chain complementarity-determining region 3 (CDR3) in patients with TAK. Five untreated patients with TAK were collected from June 2019 to December 2019. Four healthy blood samples were matched as the control group. The blood mononuclear cells were separated, and RNA was extracted for reverse transcription to obtain complementary DNA. Then high-throughput sequencing was performed. The quality of samples was evaluated by principal component analysis. We compared the diversity and expression of TCR α-chain between TAK group and control group. R software was used for statistical analysis and drawing, and Mann-Whitney U test was used to analyze the differences between the two groups. The results showed that there was a significant difference in the diversity of TCR α-chain CDR3 between the two groups. Three V region genes expression significantly higher in the TAK patients than in the control group. A total of 196 VJ rearrangement genes are significantly different between the two groups, of which 149 rearrangement genes in the TAK group are lower than those in the control group, and 47 rearrangement genes in the TAK group are higher than those in the control group. Patients with TAK have a unique TCR α-chain CDR3 library. These characteristic genes may be a marker for early diagnosis and provide a new theoretical basis for treating TAK.

Blood Collection Workflow for Interrogation of Translational Samples with Single Cell Technologies

Blood is a valuable and easily accessible sample type for clinical researchers that provides a snapshot of the organ systems at the time of sample collection. Readouts obtained from blood help inform clinical treatments, investigate disease mechanisms, evaluate drug response, and monitor outcomes. However, the impact of existing blood collection workflows on data output from new and high resolution technologies, like single cell RNA sequencing (scRNA-seq) remains a challenge. Recent scRNA-seq studies have shown that changes in gene expression can occur in blood cells in as little as a few hours post collection1. While bulk measurements rely on sample fixation to preserve the sample during transport, this approach does not preserve intact cells and hence, is not compatible with single cell technologies. With the recent availability of fixation compatible scRNA-seq assays comes the need to preserve blood while retaining single cell information. In this study, we evaluated various PBMC isolation and blood preservation methods which preserve single cell information while easing sample logistical constraints. Our initial experiments compared various PBMC isolation methods. scRNA seq analysis of these samples demonstrated that all isolation methods captured single cell gene expression with some variability in cell types along with user-variability. Changes in cell type proportions were observed in as little as 4 hrs post blood draw, emphasizing the need for an effective blood preservation method to retain single cells during transport and blood cell isolation. Next, we compared different methods of blood preservation including cryopreservation and fixation. Healthy and diseased whole blood samples were collected, preserved, stored followed by isolation of blood samples at different time points. scRNA-seq analysis of the samples demonstrated that blood can be preserved, transported, and stored (weeks to months) before blood cell isolation. Blood cells isolated from these cells can then be stored long term for scRNA-seq analysis. This blood collection and preparation workflow also eases blood transportation logistical constraints, allowing for batch shipping of samples from distributed collection sites. Coupled with automatable cell isolation methods and multiplexing single cell sequencing solutions, these workflows can be adapted for large scale translational research studies.