Whole Blood Of Patients Research Articles

Epigenome-wide DNA methylation study of whole blood in patients with sporadic amyotrophic lateral sclerosis.

Background:Epigenetics, and especially DNA methylation, contributes to the pathogenesis of sporadic amyotrophic lateral sclerosis (SALS). This study aimed to investigate the role of DNA methylation in SALS using whole blood of SALS patients.Methods:In total, 32 SALS patients and 32 healthy controls were enrolled in this study. DNA was isolated from whole blood collected from the participants. DNA methylation profiles were generated using Infinium MethylationEPIC BeadChip.Results:We identified 34 significant differentially methylated positions (DMPs) in whole blood from SALS patients, compared with the healthy controls. Of these DMPs, five were hypermethylated and 29 were hypomethylated; they corresponded to 13 genes. For the DMPs, ATAD3B and BLK were hypermethylated, whereas DDO, IQCE, ABCB1, DNAH9, FIGN, NRP1, TMEM87B, CCSAP, ST6GALNAC5, MYOM2, and RUSC1-AS1 were hypomethylated. We also identified 12 differentially methylated regions (DMRs), related to 12 genes (NWD1, LDHD, CIS, IQCE, TNF, PDE1C, LGALS1, CSNK1E, LRRC23, ENO2, ELOVL2, and ELOVL2-AS1). According to data from the Kyoto Encyclopedia of Genes and Genomes database, DNAH9 and TNF are involved in the amyotrophic lateral sclerosis (ALS) pathway. Correlation analysis between clinical features and DNA methylation profiling indicated that the methylation level of ELOVL2 and ARID1B was positively associated with the age of onset (r = 0.86, adjust P = 0.001) and disease duration (r = 0.83, adjust P = 0.01), respectively.Conclusions:We found aberrant methylation in DMP- and DMR-related genes, implying that many epigenetic alterations, such as the hypomethylation of DNAH9 and TNF, play important roles in ALS etiology. These findings can be helpful for developing new therapeutic interventions.

A Capillary Electrophoresis-Based Method for the Measurement of Hydroxychloroquine and Its Active Metabolite Desethyl Hydroxychloroquine in Whole Blood in Patients with Rheumatoid Arthritis.

A capillary electrophoresis method was developed to detect and measure hydroxychloroquine (HCQ) and its active metabolite desethyl hydroxychloroquine (DHCQ) in whole blood in patients with rheumatoid arthritis. The best separation in terms of peak area reproducibility, migration time, peak shape, and resolution of adjacent peaks was obtained in a 60 cm, 75 µm i.d. uncoated fused-silica capillary using a background electrolyte mixture of an aqueous 55 mmol/L TRIS solution brought to pH 2.6 with phosphoric acid and methanol (85:15) and a voltage and a temperature of separation of 20 kV and 30 °C, respectively. Analytes were separated in less than 12 min, with excellent linearity (R2 ≥ 0.999) in the concentration range of 0.5–8 µmol/L. The recovery of analytes spiked in whole blood was 99–101% for HCQ and 98–99% for DHCQ. Analysis of five samples from patients with rheumatoid arthritis receiving HCQ 400 mg daily yielded mean steady-state concentrations of 2.27 ± 1.61 and 1.54 ± 0.55 μmol/L for HCQ and DHCQ, respectively, with a HCQ to DHCQ ratio of 1.40 ± 0.77.

Model-informed drug development of voxelotor in sickle cell disease: Population pharmacokinetics in whole blood and plasma.

Oxbryta (voxelotor) is a small‐molecule inhibitor of sickle hemoglobin (Hb) polymerization approved for patients with sickle cell disease (SCD) aged greater than or equal to 12 years at a dose of 1500 mg once daily (q.d.). Voxelotor binds preferentially to Hb, and voxelotor partitioning into red blood cells is an effective predictor of Hb occupancy. The objectives of these analyses were to develop a population pharmacokinetic (PopPK) model for voxelotor in both plasma and whole blood in adults and adolescents to support the dose selection for optimal target engagement and to identify covariates that have a significant effect on voxelotor pharmacokinetics (PK) in plasma and whole blood. An integrated plasma and whole blood PopPK model with two compartments, first‐order absorption and elimination, and a site‐of‐action effect compartment adequately described the concentration‐time profiles of voxelotor in plasma and whole blood in patients treated up to 72 weeks. Covariates with significant effects on voxelotor PK included baseline blood volume on apparent volume of the central compartment and time‐varying hematocrit and dose on whole blood partitioning, indicating that clinical markers of voxelotor effect can, in turn, influence its PK. Furthermore, the model confirmed that voxelotor PK in plasma and whole blood is linear with dose and time and comparable for adults and adolescents. No clinically important covariate effects on voxelotor PK that warranted dose adjustment were identified in this analysis. Overall, the PopPK analyses contributed significantly to the voxelotor label and support 1500 mg q.d. as the therapeutic dose in adults and adolescents with SCD.

Identification of Regulatory Factors and Prognostic Markers in Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive degeneration of motor neurons, leading to muscle atrophy, paralysis and even death. Immune disorder, redox imbalance, autophagy disorder, and iron homeostasis disorder have been shown to play critical roles in the pathogenesis of ALS. However, the exact pathogenic genes and the underlying mechanism of ALS remain unclear. The purpose of this study was to screen for pathogenic regulatory genes and prognostic markers in ALS using bioinformatics methods. We used Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, gene set enrichment analysis (GSEA), and expression regulation network analysis to investigate the function of differentially expressed genes in the nerve tissue, lymphoid tissue, and whole blood of patients with ALS. Our results showed that the up-regulated genes were mainly involved in immune regulation and inflammation, and the down-regulated genes were mainly involved in energy metabolism and redox processes. Eleven up-regulated transcription factors (CEBPB, CEBPD, STAT5A, STAT6, RUNX1, REL, SMAD3, GABPB2, FOXO1, PAX6, and FOXJ1) and one down-regulated transcription factor (NOG) in the nerve tissue of patients with ALS likely play important regulatory roles in the pathogenesis of ALS. Based on construction and evaluation of the ALS biomarker screening model, cluster analysis of the identified characteristic genes, univariate Cox proportional hazards regression analysis, and the random survival forest algorithm, we found that MAEA, TPST1, IFNGR2, and ALAS2 may be prognostic markers regarding the survival of ALS patients. High expression of MAEA, TPST1, and IFNGR2 and low expression of ALAS2 in ALS patients may be closely related to short survival of ALS patients. Taken together, our results indicate that immune disorders, inflammation, energy metabolism, and redox imbalance may be the important pathogenic factors of ALS. CEBPB, CEBPD, STAT5A, STAT6, RUNX1, REL, SMAD3, GABPB2, FOXO1, PAX6, FOXJ1, and NOG may be important regulatory factors linked to the pathogenesis of ALS. MAEA, TPST1, IFNGR2, and ALAS2 are potential important ALS prognostic markers. Our findings provide evidence on the pathogenesis of ALS, potential targets for the development of new drugs for ALS, and important markers for predicting ALS prognosis.

The National Blood Shortage-An Impetus for Change.

The National Blood Shortage-An Impetus for Change.

Interleukin 8 in postoperative delirium – Preliminary findings from two studies

ObjectiveStudies have suggested that inflammation contributes to the pathogenesis of postoperative delirium, but previous results on the proinflammatory cytokine IL-8 in plasma are contradictory. Additionally, a significant fraction of IL-8 is bound to erythrocytes, but the relevance of whole blood IL-8 in delirium has not been studied. In this work, we analyzed the association of postoperative delirium with levels of unbound IL-8 in plasma and levels of IL-8 in whole blood in patients from two studies which were conducted in our department and have not been presented previously. We assessed the prognostic value of whole blood IL-8. MethodsPlasma/whole blood IL-8 was measured at least once in N ​= ​504 patients preoperatively, on day one (d1) and/or three months after surgery in the BioCog observational study. Whole blood IL-8 was measured in N ​= ​64 patients from the PHYDELIO trial preoperatively, on d1 and d7 after surgery. For the determination of whole blood IL-8, EDTA-preserved blood samples underwent lysis by adding Triton-X100 surfactant. Plasma and whole blood IL-8 levels were assessed with two different immunoassay kits. Delirium was appraised systematically for seven postoperative days according to DSM criteria using two comparable protocols consisting of validated screening tools. ResultsDelirium occurred in 25% of BioCog and 14% of PHYDELIO patients. In BioCog, IL-8 was elevated on d1 and in delirious patients. A steeper postoperative increase in delirium was confounded by surgery-related factors. A crescendo-decrescendo pattern of whole blood IL-8 levels was observed in non-delirious patients with a peak on d1. This pattern was more distinct in delirious BioCog patients, but inverted in delirious PHYDELIO patients. Preoperative whole blood IL-8>318.4 ​pg/mL (reference <150 ​pg/mL) had adequate sensitivity (0.79/0.78) and specificity (0.53/0.67) for delirium in both samples. ConclusionOur results contribute to an inflammatory hypothesis of postoperative delirium.

The functional consequence of the pathogenic variant in the inflammasome component NLRP3 as the underlying cause of keratitis fugax hereditaria

AbstractPurposeKeratitis fugax hereditaria (KFH) is a rare periodic autoinflammatory corneal disease caused by a heterozygous pathogenic variant c.61G&gt;C in the NLRP3 gene. Other NLRP3 variants cause cryopyrin‐associated periodic syndromes (CAPS), characterized by enhanced production of IL‐1β. While more than 50% of KFH patients use prednisolone to relieve pain, there are no data on how the c.61G&gt;C NLRP3 variant or prednisolone treatment affects IL‐1β production in KFH patients. We carried out a functional study to elucidate these questions.MethodsPeripheral blood mononuclear cells (PBMCs) isolated from whole blood of patients with KFH (n = 8) and healthy controls (n = 4) were grown under steady‐state conditions or exposed to NLRP3 inflammasome priming with LPS (500 ng/ml) together with or without prednisolone (100 µM) and subsequently activating inflammasome with ATP(1 mM). Proteins precipitated from the cell culture media were separated by SDS‐PAGE and transferred to a membrane by western blotting. IL‐1β was visualized from the membranes by immunodetection. Alternatively, the concentration of secreted IL‐1β in the media was quantified by ELISA.ResultsFollowing NLRP3 inflammasome activating stimuli, a reduced threshold for IL‐1β secretion was observed in severely affected KFH patients compared to mildly affected patients or healthy controls. In these patients, LPS priming alone was able to trigger IL‐1β secretion while in the mildly affected patients and healthy controls IL‐1β secretion required a canonical two‐hit activation of the NLRP3 inflammasome by both LPS and ATP. In both KFH and healthy control‐derived PBMCs, IL‐1β secretion declined after prednisolone treatment.ConclusionsOur study demonstrated that patients with severe KFH are more prone to IL‐1β secretion, thus indicating that the c.61G&gt;C is a gain‐of‐function variant. Furthermore, prednisolone is a potent treatment for reducing IL‐1β secretion in KFH.

Expression of miR-29a in whole Blood of Patients with Colorectal Neoplasm

Expression of miR-29a in whole Blood of Patients with Colorectal Neoplasm

Platelet Defects in Acute Myeloid Leukemia-Potential for Hemorrhagic Events.

Background and objectives: In acute myeloid leukemia (AML), extensive bleeding is one of the most frequent causes of death. Impaired activation and aggregation processes were identified in previous studies on platelet behaviour associated with this disease. This study’s aim was to examine platelet function in correlation with other haemorrhage risk factors (fever, sepsis, recent bleeding, uraemia, leucocytosis, haematocrit value, treatment). Design and methods: The analysis of platelet surface proteins (Glycoprotein Ib-IX (CD42b, CD42a), Glycoprotein IIb-IIIa (CD41, CD61), p-selectin (CD62P), granulophysin (CD63)) was conducted by flowcytometry from samples of whole blood in patients with acute myeloid leukaemia in different stages of diagnosis and therapy (n = 22) in comparison with healthy human controls (n = 10). Results and interpretations: Our results show a significant decrease in fluorescence level associated with platelet activation markers (CD63 (14.11% vs. 40.78 % p < 0.05); CD62P (15.26% vs. 28.23% p < 0.05)); adhesion markers (CD42b (69.08% vs. 84.41% p < 0.05)) and aggregation markers (CD61 (83.79% vs. 98.62% p < 0.001)) in patients compared to controls. The levels of CD41 (80.62% vs. 86.31%, p = 0.290) and CD42a (77.98% vs. 94.15%, p = 0.99) demonstrate no significant differences in the two groups. Conclusion: The AML patients present changes in adhesion receptors and activation markers, suggesting a functional defect or denatured intracellular signalling in platelets. The exposed data indicate that flow cytometry can effectively identify multiple functional platelet impairments in AML pathogenesis.

Using Bioinformatics and NCBI Tools to Sequence and Structure Analysis of Transcription Factor 7 Like 2 Gene (TCF7L2) in Iraqi Diabetic Mellitus Type II Patients

In this paper, the study aimed to analyze the sequence and structure of Transcription Factor 7 Like 2 gene (TCF7L2) in Iraqi Diabetic Mellitus Type II (T2DM) Patients and was compared with standard sequence form National Center of Bioinformatics Tools (NCBI) using BLAST. Ten blood samples of (T2DM) Iraqi patients, was collected from Al-Mustansiriya University National Diabetes Centre in Baghdad Province/Iraq (17-65 year). DNA was extracted from whole blood of patients’ samples using the Quick-DNA™ Blood MiniPrep kit, then it was sent to Korea at Macrogen Corporartion Company, where they used automated DNA sequencing for sequence analysis. The results of the sequence analysis of (T2DM) patients’ samples, we found: six missense mutations, one deletion mutation, and three silent mutations detected using BLAST in (NCBI). All mutations appeared at the same sites of the gene which controls the rate of genetic information transcription that indicates to have a relationship with (T2DM). These mutations were recorded on the (NCBI). The physicochemical properties of (TCF7L2) determined in the present study included; alpha-helical structure and 3-Dimension structure appeared contrast when compared with the gene template. Briefly, mutations effected (TCF7L2) which influences the structure, physicochemical properties of the protein, and the secretion of insulin hormone which maintains glucose level in blood.

Clinical significance of acetylcholinesterase activity in brain tumors.

It has been established that the non-neuronal cholinergic system is related to the oncogenesis which increases the attractiveness of its components as the promising markers of oncologic diseases. The purpose of this work is to evaluate the clinical significance of the analysis of the activity of acetyl cholinesterase as a new marker of gliomas. The activity of acetyl cholinesterase was assessed by photo colorimetric analysis according to the Hestrin method recalculating the activity of the enzyme in the tumor tissue per 1 g of protein, and in the blood - by 0.1 g of hemoglobin. The data obtained in the primary tumors of the brain (28) in the tissue of the brain of persons who died as a result of injury (6) and in whole blood of patients with gliomas (28) and practically healthy people (10) were compared with the use of a number of statistical programs. A significant decrease in the activity of acetyl cholinesterase in tumor tissue and in whole blood is revealed as the degree of anaplasia of tumors increases, starting with Grade II. It is for the first time that a significant direct correlation was noted showing the consistency between the decrease in the activity of acetyl cholinesterase in the tumor tissue of the brain and blood. Bioinformatic analysis showed the connection of the enzyme of acetyl cholinesterase with proteins of the PI3K-AKT and Notch signaling pathways providing antiapoptotic and proliferative effects. The found dependences provide new insights into understanding of the mechanisms of gliomas genesis and can be used for selection of new diagnostic markers of brain tumors.

Integrated Gene Expression Profiling Analysis Reveals Potential Molecular Mechanisms and Candidate Biomarkers for Early Risk Stratification and Prediction of STEMI and Post-STEMI Heart Failure Patients.

Objective: To explore the molecular mechanism and search for the candidate differentially expressed genes (DEGs) with the predictive and prognostic potentiality that is detectable in the whole blood of patients with ST-segment elevation (STEMI) and those with post-STEMI HF.Methods: In this study, we downloaded GSE60993, GSE61144, GSE66360, and GSE59867 datasets from the NCBI-GEO database. DEGs of the datasets were investigated using R. Gene ontology (GO) and pathway enrichment were performed via ClueGO, CluePedia, and DAVID database. A protein interaction network was constructed via STRING. Enriched hub genes were analyzed by Cytoscape software. The least absolute shrinkage and selection operator (LASSO) logistic regression algorithm and receiver operating characteristics analyses were performed to build machine learning models for predicting STEMI. Hub genes for further validated in patients with post-STEMI HF from GSE59867.Results: We identified 90 upregulated DEGs and nine downregulated DEGs convergence in the three datasets (|log2FC| ≥ 0.8 and adjusted p < 0.05). They were mainly enriched in GO terms relating to cytokine secretion, pattern recognition receptors signaling pathway, and immune cells activation. A cluster of eight genes including ITGAM, CLEC4D, SLC2A3, BST1, MCEMP1, PLAUR, GPR97, and MMP25 was found to be significant. A machine learning model built by SLC2A3, CLEC4D, GPR97, PLAUR, and BST1 exerted great value for STEMI prediction. Besides, ITGAM and BST1 might be candidate prognostic DEGs for post-STEMI HF.Conclusions: We reanalyzed the integrated transcriptomic signature of patients with STEMI showing predictive potentiality and revealed new insights and specific prospective DEGs for STEMI risk stratification and HF development.

Laboratory-based dried blood spot testing for hepatitis C: A protocol for systematic review and meta-analysis of diagnostic accuracy

Background: Diagnosis of chronic hepatitis C virus (HCV) infection typically involves collection of venous blood samples prior to serological investigation of an antibody response followed by a confirmatory viral load or antigen test to verify active HCV infection. This conventional pathway poses logistical challenges for the implementation of reflex testing, whereby the confirmatory test is performed on the same sample used for serological investigation. Dried blood spot (DBS) testing, in which capillary blood is deposited on filter paper, is a less invasive alternative that can enable reflex testing without the need for venepuncture, centrifugation and freezing of samples. Methods: This systematic review aims to assess the diagnostic accuracy of DBS compared with venous blood samples for diagnosis of chronic HCV infection. Observational studies which compare diagnostic tests using DBS with those using serum, plasma or whole blood in patients with chronic or resolved HCV infection will be included. Electronic searches will be conducted in PubMed, Embase, Scopus, Web of Science, Lilacs and the Cochrane library. Citation screening, data extraction and quality appraisal of included studies will be performed in duplicate using the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) tool. A meta-analysis will be conducted to derive pooled estimates of sensitivity, specificity, positive likelihood ratios, negative likelihood ratios, and diagnostic odds ratios. Sensitivity analyses and meta-regression will also be performed. Quality of the evidence will be evaluated using the GRADE criteria. Discussion: Identifying and linking people with currently undiagnosed chronic HCV infection to care is pivotal to attaining global viral hepatitis elimination targets. The use of DBS could simplify diagnostic testing strategies by integrating reflex testing into the care pathway and reducing drop-off along the cascade of care. Registration: PROSPERO, CRD42020205204. Registered 19 th September 2020.

Expression of Microrna-195 Increased Significantly in Patients with Methamphetamine Abuse Disorder

Background: Addiction is one of the most important social and health problems in the world. Development of rapid and inexpensive diagnostic methods for identification of patients with addiction to methamphetamine is still a very important challenge. Recently, microRNAs (miRNAs) have been introduced as an accurate and reliable biomarker for diagnosis of human disorders. Objectives: In the present study, the expression of miRNA-186 and miRNA-195 was investigated in blood of patients with methamphetamine abuse disorder. Methods: In this case-control study, 60 patients with methamphetamine abuse disorder (case group) and 60 healthy controls (control group) were enrolled. Total RNA was extracted from whole blood of patients and healthy controls, and then cDNA synthesis was performed using reverse transcriptase. Real-time PCR method was employed to investigate miRNA-186 and miRNA-195 expression. Finally, statistical software was used to analyze the obtained data. Results: The results demonstrated that the expression of miRNA-195 significantly increased in blood samples of patients with methamphetamine abuse disorders (8.75-fold change) compared to healthy controls (P &lt; 0.05). However, the expression of miRNA-185 did not significantly increase (1.61-fold change) in patients compared to healthy controls (P &gt; 0.05). Conclusions: Our study suggested that miR-195 may play an important role in the pathogenesis of drug addiction and can be used as an accurate and reliable marker for the identification of patients with methamphetamine abuse disorder.

Circulating circTOLLIP serves as a diagnostic biomarker for liquid biopsy in non-small cell lung cancer

BackgroundCircular RNAs (CircRNAs) have been found to possess vital functions in tumorigenesis of various cancer types, including non-small cell lung cancer (NSCLC). The aim of this study was to identify and explore the diagnostic values of the newly found Toll interacting protein (TOLLIP)-derived circRNA (circTOLLIP) for liquid biopsy in NSCLC. MethodsRNase R and actinomycin D assays were conducted to confirm the existence and stability of circTOLLIP. RT-qPCR was performed to identify the expression levels of circTOLLIP in NSCLC tumor tissues, whole blood, and cell lines. The diagnostic values were evaluated by receiver operating characteristic (ROC) curve analysis. ResultsCircTOLLIP was screened as a candidate biomarker and was found to be significantly down-regulated in both NSCLC tissues and cell lines. Interestingly, circulating circTOLLIP was also lower-expressed in the whole blood of patients with NSCLC compared to that of patients with benign lung disease and healthy controls. Importantly, the circulating circTOLLIP represented better diagnostic values in comparison to the traditional tumor markers (NSE, CYFR21-1, and CA72-4), and showed higher stability even though the whole blood was exposed to various tough conditions. ConclusionsOur findings indicate that circTOLLIP can be used as a non-invasive biomarker to distinguish early-stage NSCLC from benign lung diseases and from healthy controls, suggesting the potential application of circTOLLIP for liquid biopsy in NSCLC.

Genome-Wide DNA Methylation Profile in Whole Blood of Patients With Chronic Spontaneous Urticaria.

BackgroundChronic spontaneous urticaria (CSU) is a common autoimmune skin disease. Little is known about the role of epigenetics in the pathogenesis of CSU. This study aimed to investigate genome-wide DNA methylation profile in whole blood of patients with CSU.Patients and MethodsGenome-wide DNA methylation levels in whole blood samples of 95 Chinese Han ethnicity adult CSU patients and 95 ethnicity-, age- and sex-matched healthy controls were analyzed using Illumina 850K methylation chip. The differentially methylated genes (DMGs) were screened out and then functionally annotated by the gene ontology and the Kyoto encyclopedia of genes and genomes databases.ResultsA total of 439 differentially methylated positions (DMPs) (p < 0.01 and |Δβ| ≥ 0.06) were identified with 380 hypomethylated and 59 hypermethylated. The average global DNA methylation levels of the 439 DMPs in the CSU patients were significantly lower than those in the healthy controls (p < 0.001). The distribution of the 439 DMPs was wide on chromosome 1 to 22 and chromosome X. Chromosome 6 embodied the largest number of DMPs (n = 51) and their annotated genes were predominantly related to autoimmunity. The 304 annotated DMGs were mainly enriched in autoimmune disease- and immune-related pathways. A total of 41 DMPs annotated to 28 DMGs were identified when p < 0.01 and |Δβ| ≥ 0.1. Of the 28 DMGs, HLA-DPB2, HLA-DRB1, PPP2R5C, and LTF were associated with autoimmunity. CSU cases with elevated total IgE, positive anti-thyroid peroxidase IgG autoantibodies, positive anti-thyroglobulin IgG autoantibodies, angioedema, UASday > 4, or recurrent CSU showed phenotype-specific DMPs as compared with cases with normal total IgE, negative anti-thyroid peroxidase IgG autoantibodies, negative anti-thyroglobulin IgG autoantibodies, no angioedema, UASday ≤ 4, or non-recurrent CSU respectively.ConclusionThis study shows a distinct genome-wide DNA methylation profile in Chinese Han ethnicity adult CSU patients and indicates a role of epigenetics in the pathogenesis of CSU. The predominant enrichment of the CSU-associated DMGs in immunological pathways provides supportive evidence for the immunopathogenesis of CSU. Future research on the CSU-associated DMPs and DMGs will help discover potential therapeutic targets for CSU.

Whole blood gene expression signature in patients with obstructive sleep apnea and effect of continuous positive airway pressure treatment

The molecular mechanisms of obstructive sleep apnea (OSA), in particular the gene expression patterns in whole blood of patients with OSA, can shed more light on the underlying pathophysiology of OSA and suggest potential biomarkers. In the current study, we have enrolled thirty patients with untreated moderate-severe OSA together with 20 BMI, age, and sex-matched controls and 15 normal-weight controls. RNA-sequencing of whole blood and home sleep apnea testing were performed in the untreated state and after three and twelve months of continuous positive airway pressure (CPAP) treatment. Analysis of the whole blood transcriptome of the patients with OSA revealed a unique pattern of differential expression with a significant number of downregulated immune-related genes including many heavy and light chain immunoglobulins and interferon-inducible genes. This was confirmed by the gene ontology analysis demonstrating enrichment with the biological processes associated with various immune functions. Expression of these genes was recovered after three months of CPAP treatment. After 12 months of CPAP treatment, the overall gene expression profile returns to the initial, untreated level. In addition, we have confirmed the importance of choosing BMI-matched controls as a reference group as opposed to normal-weight healthy individuals based on the significantly different gene expression signatures between these two groups.

Expression Pattern of Leptin and Its Receptors in Endometrioid Endometrial Cancer.

The identification of novel molecular markers and the development of cancer treatment strategies are very important as cancer incidence is still very high. Obesity can contribute to cancer progression, including endometrial cancer. Adipocytes secrete leptin, which, when at a high level, is associated with an increased risk of cancer. The aim of this study was to determine the expression profile of leptin-related genes in the endometrial tissue samples and whole blood of patients. The study material included tissue samples and whole blood collected from 30 patients with endometrial cancer and 30 without cancer. Microarrays were used to assess the expression profile of leptin-related genes. Then, the expression of leptin (LEP), leptin receptor (LEPR), leptin receptor overlapping transcript (LEPROT), and leptin receptor overlapping transcript-like 1 (LEPROTL1) was determined by the Real-Time Quantitative Reverse Transcription Reaction (RT-qPCR). The serum leptin concentration was evaluated using Enzyme-linked immunosorbent assay (ELISA). Leptin and its receptors were overexpressed both at the mRNA and protein levels. Furthermore, there were strong positive correlations between leptin levels and patient Body Mass Index (BMI). Elevated levels of leptin and its receptors may potentially contribute to the progression of endometrial cancer. These observations may be useful in designing endometrial cancer treatment strategies.

МИНИ-ФОТОФЕРЕЗ – ЭКСТРАКОРПОРАЛЬНЫЙ ФОТОФЕРЕЗ, НЕ ТРЕБУЮЩИЙ ЛЕЙКАФЕРЕЗА: ОПЫТ КЛИНИЧЕСКОГО ИСПОЛЬЗОВАНИЯ

Extracorporeal photopheresis (ECP) has proven effectiveness for treatment of several diseases, including acute and chronic graft-versus-host disease (GVHD) after allogenic transplantation of hematopoietic blood stem cells. The standard ECP requires leukapheresis to obtain a mononuclear cell fraction. The possibility of using leukapheresis is limited by the requirements for vascular access and the somatic status of the patient. There is a relatively new method of performing ECF, called «mini-photopheresis» (mini-ECF), in which a fraction of mononuclear cells is isolated from a dose of whole blood obtained by the exfusion method. The article presents preliminary results of using mini-ECP in patients with acute and chronic GVHD. Materials and methods of research: the study included 11 patients with acute (7 patients) and chronic (4 patients) GVHD who received mini-ECP therapy from June 2018 to January 2021. Leukocyte fractions rich in mononuclear cells were prepared from the dose of whole blood of patients. The resulting fraction was diluted with 0,9% NaCl solution to less than 3% hematocrit. The cellular product was then injected with an 8-Methoxyperalene and programmed with UV spectrum A. Autologous erythrocytes and the finished cellular product were injected into the patient after irradiation. Results: 6 out of 7 patients (85,7%) with acute GVHD has responded to mini-ECP therapy. In patients with chronic GVHD, the response rate to mini-ECP therapy was 25%. In both groups there are no significant differences found in the number of leukocytes count per body mass in the finished cellular product. The correlation between the presence and severity of response to mini-ECP therapy with the number of leukocytes in the finished cellular product was not determined. None of the patients had adverse reactions and complications associated with mini-ECP therapy. Conclusion: mini-ECP is an attractive alternative for treatment of patients with steroid-resistant or steroid-dependent GVHD who cannot undergo leukapheresis. Our results are preliminary, but promising. We will continue to use this method as a second-line therapy for patients with contraindications to leukapheresis.

Genetic and pathophysiological substantiation of polyphenolic grape processing products’ application in the treatment of metabolic syndrome in the population of the Republic of Crimea

Adipose tissue in abdominal obesity produces various cytokines, the most important of which is adiponectin (AN). Polymorphism of the AN receptor genes is associated with the risk of metabolic syndrome (MS) and type 2 diabetes mellitus (DM2) development. The aim of the study was to investigate the association of AN genes receptors’ polymorphism with the development of MS and DM2 in the Republic of Crimea as well as the possibility of named pathology correction with polyphenolic functional foods. The study included 100 patients with confirmed diagnosis of DM2 and MS. Genomic DNA was isolated from the whole blood of patients. Gene polymorphism amplification was performed by PCR real-time. The most common genotype of the ADIPOQ gene (rs1501299) among patients with DM2 and MS was GT polymorphism G276T. The most frequent genotypes of the ADIPOR1 gene were AC (rs2275737) – 53.3% and CT (rs2275738) - 50.0%. The most frequent combinations between ADIPOR1 and ADIPOQ genes were AC (rs2275737) /CT (rs2275738) and GT (rs1501299) – 28.6%, as well as AC (rs2275737)/CT (rs2275738) and GG (rs1501299) – 19.7%. Thus, in patients with MS and DM2 in the Republic of Crimea, associations of polymorphism of the genes of the AN receptors with the development of this pathology have been established. It is known that polyphenols of red grape varieties are epigenetic modifiers. Therefore, these products can be included in individual nutrition programs in population groups genetically predisposed to MS and DM2.