Elevated levels of Th17 cells in children with central obesity

<div><p></p><p><b><i>Background</i>.</b> It is believed that the recently discovered interleukin 17-producing Th17 cells play a role in the pathogenesis of chronic inflammation in the course of obesity and diabetes. <b><i>Objectives</i>.</b> The purpose of our study was to complete data on this subject in children. <b><i>Methods</i>.</b> We assessed Th17 cell levels in the peripheral blood of children diagnosed with central obesity (<i>n</i> = 14) and compared the results with data obtained in patients with newly diagnosed (<i>n</i> = 11) and long-term type 1 diabetes mellitus (<i>n</i> = 18), and in a control group as well (<i>n</i> = 24). <b><i>Results</i>.</b> (i) Children with central obesity were characterized by higher percentages of Th17 cells as compared to children from the control group; (ii) in the peripheral blood of patients with long-term type 1 diabetes the Th17 cell counts were higher compared to the control group; (iii) total plasma cholesterol concentration correlated positively with Th17/Treg cells ratio; and (iv) among patients with long-term diabetes, disease duration correlated positively with Th17 cell count and Th17/Th1 cell ratio. <b><i>Conclusion</i>.</b> The results of our study indicate that Th17 cells may be involved in chronic inflammation accompanying obesity and type 1 diabetes mellitus in children.</p></div

Lower proportion of CD19⁺IL-10⁺ and CD19⁺CD24⁺CD27⁺ but not CD1d⁺CD5⁺CD19⁺CD24⁺CD27⁺ IL-10⁺ B cells in children with autoimmune thyroid diseases

Introduction: As it is generally known, regulatory B cells (Bregs) control inflammation and autoimmunity. The significance of Bregs in the population of children with autoimmune thyroid diseases (AITD) still offers plenty of potential to explore. The aim of this study was to estimate the expression of Bregs (phenotype CD19+CD24+CD27+IL-10+, CD19+IL-10+, CD1d+CD5+CD19+IL-10+ and CD1d+CD5+CD19+CD24+CD27+) in a paediatric cohort with AITD and in health controls. Materials and methods: A total of 100 blood samples were obtained from 53 paediatric patients with Graves’ disease (GD) (N = 12 newly diagnosed, mean age 12.5 ± 3.5 and N = 17 during methimazole therapy, mean age 12.7 ± 4.4), Hashimoto’s thyroiditis (HT) (N = 10 newly diagnosed, mean age 13.3 ± 2.9 and N = 10 during L-thyroxine therapy, mean age 13.7 ± 3.4) and compared with healthy controls (C) (N = 15, mean age 13.1 ± 3.1). The expressions of the immune cell populations were analysed by four-color flow cytometry using a FASC Canto II cytometer (BD Biosciences). Results: There was a decreasing tendency in the number of lymphocytes B producing IL-10 (B10) cells among all B lymphocytes and more widely, also among all lymphocytes, in each study group, as compared to C. We reported a reduction in IL-10 production in Bregs with the expression of CD19+CD24+CD27+IL-10 and CD1d+CD5+CD19+IL-10+ in both untreated and treated AITD. Conclusions: Our data demonstrate that the reduction in the number of Bregs with CD19+CD24+CD27+IL-10+ and CD19+IL-10+ expression could be responsible for breaking immune tolerance and for AITD development in children.</p

DataSheet_1_Effect of antiviral and immunomodulatory treatment on a cytokine profile in patients with COVID-19.docx

BackgroundThe severity of COVID-19 is associated with an elevated level of a variety of inflammatory mediators. Increasing evidence suggests that the Th17 response contributes to the severity of COVID-19 pneumonia, whereas Th22 response plays a regulatory role in SARS-CoV-2 infection. Two main types of available COVID-19 treatments are antivirals and immunomodulatory drugs; however, their effect on a cytokine profile is yet to be determined.MethodsThis study aim to analyse a cytokine profile in peripheral blood from patients with COVID-19 (n=44) undergoing antiviral or/and immunomodulatory treatment and healthy controls (n=20). Circulating CD4+ and CD8+ T cells and their intracellular expression of IL-17A and IL-22 were assessed by flow cytometry.ResultsInitial results showed an overexpression of IL-17F, IL-17A, CCL5/RANTES, GM-CSF, IL-4, IL-10, CXCL-10/IP-10 and IL-6 in COVID-19 patients compared to healthy controls. Treatment with remdesivir resulted in a significant decline in concentrations of IL-6, IL-10, IFN-alpha and CXCL10/IP-10. Immunomodulatory treatment contributed to a significant downregulation of IL-10, IFN-alpha, CXCL10/IP-10 and B7-H3 as well as upregulation of IL-22 and IL-1 beta. A combination of an antiviral and immunomodulatory treatment resulted in a significant decrease in IL-17F, IL-10, IFN-alpha, CXCL10/IP-10 and B7-H3 levels as well as an increase in IL-17A and IL-1 beta. We found significantly higher percentage of both CD4+ and CD8+ T cells producing IL-17A and CD4+ T cells producing IL-22 in patients with COVID-19.ConclusionAdministration of antiviral or/and immunomodulatory treatment resulted in a significant downregulation of pro-inflammatory cytokine expression and an upregulation of T cell absolute counts in most cases, thus showing effectiveness of treatment in COVID-19. SARS-CoV-2 infection induced cytokine overexpression in hospitalized patients with COVID-19 as well as lymphopenia, particularly a decrease in CD4+ and CD8+ T cell counts. Moreover, despite the reduced counts of CD4+ and CD8+ T cells, both subsets showed overactivation and increased expression of IL-17A and IL-22, thus targeting Th17 response might alleviate inflammatory response in severe disease.</p

Additional file 1: of Increased methylation upstream of the MEG3 promotor is observed in acute myeloid leukemia patients with better overall survival

Scheme of densitometry analysis. a Equal amounts of PCR product (treated and untreated with restriction enzyme, RE) were separated on a gel, and a picture of the gel was captured. Step (1) Selection of bands for analysis. Step (2) Intensity of the bands is translated to the surface of the corresponding peaks using ImageJ software. Step (3) Analysis of the surface of the peaks corresponding to particular bands and re-calculation of arbitrary units to percentages. In analysis 1, the surface of the “unmethylated” band from the re-treated sample is compared with the total amount of PCR product that was used for digestion. In analysis 2, the surface of the “unmethylated” band is compared to the total surface of all peaks in the digested sample. The mean of the methylation from both analyses is then used for further studies. Gels are shown for the COBRA analysis of CG6 (b), CG7 (c), CG8 (d), CG9 (e), ZAC (f), IGF2-H19 (g), PEG1 (h), and PEG3 (i). (PDF 1340 kb

Additional file 3: Figure S2. of Extracellular nucleotides as novel, underappreciated pro-metastatic factors that stimulate purinergic signaling in human lung cancer cells

Extracellular nucleotides induce migration and adhesion of lung cancer cell lines. Panel A. Chemotaxis of tested lung cancer cell lines in response to extracellular TTP, UTP, CTP, and GTP. Chemotaxis in response to a supraphysiological dose of HGF (10 ng/ml) was used as a control. Panel B. Adhesion of tested lung cancer cell lines to fibronectin after stimulation with extracellular TTP, UTP, CTP, and GTP. Panel C. UTP stimulates intracellular calcium release in human lung cancer cells lines. All data are shown as means ± SD with *p < 0.05. (ZIP 296 kb

Additional file 2: Figure S1. of Extracellular nucleotides as novel, underappreciated pro-metastatic factors that stimulate purinergic signaling in human lung cancer cells

Evaluation of the number of BM cells and the level of EXN in plasma after irradiation or chemotheraphy. Panel A Total number of cells isolated from tibia and femurs of animals 24 h after irradiation (0–1500 cGy) or vincristine administration (0.5–2 mg/kg). Combine results from three independent isolations. Panel B The level of ATP, UTP and adenosine in murine plasma isolated from animals 24 h after irradiation (0–1500 cGy) or vincristine administration (0.5–2 mg/kg). (PDF 229 kb

Additional file 6: Figure S5. of Extracellular nucleotides as novel, underappreciated pro-metastatic factors that stimulate purinergic signaling in human lung cancer cells

Autocrine release of ATP plays a role in migration of lung cancer cells to HGF. Chemotaxis results of HTB177 cell line to HGF (10 ng/ml) in the absence or presence of apyrase (50U/ml). (PDF 87 kb

Boletín de Segovia: Número 1 - 1837 enero 3

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200

Table1_Circulating serum miR-362-3p and miR-6721-5p as potential biomarkers for classification patients with adult-type diffuse glioma.docx

According to the fifth edition of the WHO Classification of Tumours of the Central Nervous System (CNS) published in 2021, grade 4 gliomas classification includes IDH-mutant astrocytomas and wild-type IDH glioblastomas. Unfortunately, despite precision oncology development, the prognosis for patients with grade 4 glioma remains poor, indicating an urgent need for better diagnostic and therapeutic strategies. Circulating miRNAs besides being important regulators of cancer development could serve as promising diagnostic biomarkers for patients with grade 4 glioma. Here, we propose a two-miRNA miR-362-3p and miR-6721-5p screening signature for serum for non-invasive classification of identified glioma cases into the highest-grade 4 and lower-grade gliomas. A total of 102 samples were included in this study, comprising 78 grade 4 glioma cases and 24 grade 2–3 glioma subjects. Using the NanoString platform, seven miRNAs were identified as differentially expressed (DE), which was subsequently confirmed via RT-qPCR analysis. Next, numerous combinations of DE miRNAs were employed to develop classification models. The dual panel of miR-362-3p and miR-6721-5p displayed the highest diagnostic value to differentiate grade 4 patients and lower grade cases with an AUC of 0.867. Additionally, this signature also had a high AUC = 0.854 in the verification cohorts by RT-qPCR and an AUC = 0.842 using external data from the GEO public database. The functional annotation analyses of predicted DE miRNA target genes showed their primary involvement in the STAT3 and HIF-1 signalling pathways and the signalling pathway of pluripotency of stem cells and glioblastoma-related pathways. For additional exploration of miRNA expression patterns correlated with glioma, we performed the Weighted Gene-Co Expression Network Analysis (WGCNA). We showed that the modules most associated with glioma grade contained as many as six DE miRNAs. In conclusion, this study presents the first evidence of serum miRNA expression profiling in adult-type diffuse glioma using a classification based on the WHO 2021 guidelines. We expect that the discovered dual miR-362-3p and miR-6721-5p signatures have the potential to be utilised for grading gliomas in clinical applications.</p

DataSheet1_First insight about the ability of specific glycerophospholipids to discriminate non-small cell lung cancer subtypes.xlsx

Introduction: Discrimination between adenocarcinoma (ADC) and squamous cell carcinoma (SCC) subtypes in non-small cell lung cancer (NSCLC) patients is a significant challenge in oncology. Lipidomics analysis provides a promising approach for this differentiation.Methods: In an accompanying paper, we explored oxPCs levels in a cohort of 200 NSCLC patients. In this research, we utilized liquid chromatography coupled with mass spectrometry (LC-MS) to analyze the lipidomics profile of matching tissue and plasma samples from 25 NSCLC patients, comprising 11 ADC and 14 SCC cases. This study builds upon our previous findings, which highlighted the elevation of oxidised phosphatidylcholines (oxPCs) in NSCLC patients.Results: We identified eight lipid biomarkers that effectively differentiate between ADC and SCC subtypes using an untargeted approach. Notably, we observed a significant increase in plasma LPA 20:4, LPA 18:1, and LPA 18:2 levels in the ADC group compared to the SCC group. Conversely, tumour PC 16:0/18:2, PC 16:0/4:0; CHO, and plasma PC 16:0/18:2; OH, PC 18:0/20:4; OH, PC 16:0/20:4; OOH levels were significantly higher in the ADC group.Discussion: Our study is the first to report that plasma LPA levels can distinguish between ADC and SCC patients in NSCLC, suggesting a potential role for LPAs in NSCLC subtyping. This finding warrants further investigation into the mechanisms underlying these differences and their clinical implications.</p