http://hdl.handle.net/11089/1305 Wed, 04 Nov 2026 23:34:20 GMT 2026-11-04T23:34:20Z http://hdl.handle.net/11089/59570 Platelet miRNA-Based Signature as a Potential Non-Invasive Tool for Discriminating Secondary Progressive Multiple Sclerosis (dataset) Dziedzic, Angela Background: Multiple sclerosis (MS) lacks minimally invasive biomarkers that reflect early pathogenic mechanisms. Because blood platelets contribute to throm-bo-inflammatory signaling and are major vascular carriers of regulatory miRNAs, we investigated whether platelet miRNA alterations are associated with platelet hyperre-activity in MS and whether these changes originate during thrombopoiesis by examining megakaryocytes. Methods: Platelets and megakaryocytes from MS patients and healthy controls were analyzed by transmission electron microscopy, flow cytometry, ELISA, microarray-based miRNA profiling, and Real-Time PCR validation. Logistic regression and bioinformatic analyses were used to evaluate biomarker potential and biological relevance. Results: MS platelets exhibited a hyperreactive phenotype, characterized by enhanced degranulation, microparticle release, P-selectin expression, and collagen-induced aggregation. GPVI levels were significantly elevated in platelets but remained un-changed in megakaryocytes. miRNA profiling revealed widespread downregulation in platelets, whereas megakaryocytes showed selective upregulation of miR-223-3p. A two-miRNA model (let-7a-5p and miR-146a-5p) demonstrated strong discriminatory performance (AUC = 0.89; 89% sensitivity; 85% specificity). Dysregulated miRNAs were associated with pathways involved in neuroinflammation and neurodegeneration. Conclusions: MS is associated with a distinct platelet miRNA signature linked to acquired platelet hyperreactivity and vesiculation. Platelet-derived let-7a-5p and miR-146a-5p represent promising non-invasive biomarkers supporting a mechanistic link between platelet dysfunction, vascular injury, and neuroinflammation in MS. Manuscript ID: ijms-4666041 Short description of the files: TEM_graphs_raw.pdf: Raw transmission electron microscopy (TEM) micrographs showing the ultrastructure of blood platelets from healthy controls (panels A–B) and patients with multiple sclerosis (panels C–F). Images were acquired using a JEM 1010 transmission electron microscope (JEOL Ltd., Japan) operated at 80 kV. at original acquisition magnifications of ×10, ×25, and ×40, without subsequent digital zoom. Flow cytometry dataset.xlsx: Individual patient-level data on platelet aggregates (PAGs), platelet-derived microparticles (PMPs), and P-selectin-positive platelets in healthy controls (Group 0) and multiple sclerosis patients (Group 1). The dataset includes sample identifiers (ID), group assignments, and the percentages of PAGs, PMPs, and CD62P-positive platelets measured under unstimulated conditions and following collagen stimulation (1 µg/mL). Results are expressed as percentages relative to 15,000 acquired CD61-positive events per sample. PAGs and PMPs were identified based on forward scatter (FSC) thresholds of >10^³·⁵ and <10^²·², respectively, using a logarithmic scale. Platelet P-selectin expression was determined as the percentage of CD61⁺CD62P⁺ events. All measurements were performed using standardized acquisition and gating settings on a Cube 6 flow cytometer, and data were analyzed using CyView Software v.1.5.5.8 (Sysmex Partec). GPVI level ELISA raw data.xls: Dataset containing blank-corrected absorbance measurements (OD 450 nm) and calculated glycoprotein VI (GPVI) concentrations (ng/mL) in blood platelets and megakaryocytes from patients with multiple sclerosis and healthy controls. The file includes original microplate reader outputs, sample identifiers, plate layouts, calibration data, and individual GPVI concentrations calculated using a four-parameter logistic (4PL) standard curve. Absorbance measurements were acquired using a SPECTROstar Nano microplate reader (BMG LABTECH). qPCR Raw Ct value miRNA: Dataset containing individual cycle threshold (Ct) values obtained by Real-Time PCR analysis of miRNA expression in blood platelets and megakaryocytes from patients with multiple sclerosis and healthy controls. Each worksheet corresponds to a different miRNA and includes sample identifiers, raw Ct values for the target miRNA, endogenous reference miR-191-5p, and exogenous spike-in control cel-miR-39-3p, together with calculated normalization values (ΔCt, −ΔCt) and relative expression values (2^−ΔCt). Real-Time PCR measurements were performed using a Bio-Rad CFX96 real-time PCR detection system (Bio-Rad Laboratories, USA). Microarray analysis Gene_Spring: Processed miRNA microarray data obtained from blood platelets and megakaryocytes of multiple sclerosis patients and healthy controls using the Agilent SurePrint G3 Human miRNA Microarray Release 21.0 (Design ID 070156). The dataset contains miRNA identifiers, fold changes (FC), log2 fold changes (log2FC), and expression regulation (up/down), providing differential miRNA expression profiles in both cell populations. Wed, 28 Oct 2026 00:00:00 GMT http://hdl.handle.net/11089/59570 2026-10-28T00:00:00Z http://hdl.handle.net/11089/59451 Effects of polyethylene nanoparticles on the haemostatic system in vitro: insights into coagulation cascade and fibrinolysis. Dataset. Michaś, Karolina; Liudvytska, Oleksandra; Kołodziejczyk-Czepas, Joanna Activity of Xa factor dataset. Amidolitic activity of thrombin dataset. CFF assay dataset. Clotting times dataset. Fibrinogen polymerization (fibrinogen and thrombin treated) dataset. SDS-PAGE based analysis of the plasmin-metiated fibrinogen degradation (dataset and gel images after electrophoresis). SEM images. tPa and uPa inducted plasmin activity dataset. Thu, 01 Jan 2026 00:00:00 GMT http://hdl.handle.net/11089/59451 2026-01-01T00:00:00Z http://hdl.handle.net/11089/58796 Yeast display for directed evolution of amadoriase, an enzyme capable of cleaving protein glycation products - MINIATURA 8 (2024/08/X/NZ1/01781) (dataset) Studzian, Maciej This dataset was generated as part of the Miniatura 8 project, which focused on amadoriases. These FAD-dependent enzymes catalyze the breakdown of glycated amino acids, producing a free amino acid, glucosone, and hydrogen peroxide. Naturally occurring in fungi and bacteria, they are of considerable practical importance due to their applications in medical diagnostics and their potential therapeutic use, particularly in the treatment of diseases such as diabetes. Previous attempts to improve amadoriase I from Aspergillus through targeted amino acid substitutions increased the enzyme's thermostability but did not enhance its catalytic activity. Similarly, directed evolution of the related enzyme amadoriase II resulted in only marginal improvements. The aim of this project was to develop methods for the in vitro evolution of amadoriases to increase their activity using a non-directed, ultra-high-throughput approach based on yeast surface display. This technique enables the generation of random enzyme variants displayed on the surface of yeast cells, which can then be screened for catalytic activity. A biotin- and azide-modified fructoselysine analogue, designed to mimic a glycated substrate, was successfully synthesized and immobilized on the yeast surface to serve as a substrate for activity screening. Upon catalysis by an amadoriase, the substrate was expected to lose its fluorescence, allowing fluorescence-activated cell sorting (FACS) to isolate yeast cells displaying enzyme variants with higher catalytic activity. A previously engineered variant of amadoriase II from Aspergillus fumigatus (AmaII_SII_82) was used as the starting point for the evolution experiments. Unfortunately, because the initial enzyme exhibited only negligible activity toward the biotin-modified substrate, the variants obtained after three rounds of sorting also failed to show the expected improvement in activity. Nevertheless, the techniques developed during this research provide a strong foundation for future studies aimed at improving other classes of deglycating enzymes, such as FN3K and FrlB/FrlD. Mon, 01 Jun 2026 00:00:00 GMT http://hdl.handle.net/11089/58796 2026-06-01T00:00:00Z http://hdl.handle.net/11089/58757 Ochronna rola CORMs w łagodzeniu stresu oksydacyjnego i stanu zapalnego indukowanego doksorubicyną w modelu prawidłowych komórek skóry - MINIATURA 8 2024/08/X/NZ7/01320 (dataset) Juszczak, Michał Doxorubicin is a potent anticancer drug that unfortunately damages healthy cells. Researchers from the University of Łódź investigated whether carbon monoxide could protect against these side effects. They used CORMs (carbon monoxide-releasing molecules)—specifically CORM-2 and CORM-3—which safely deliver the gas to cells. The team tested these on healthy human fibroblasts exposed to doxorubicin, alongside inactive, gas-free molecules as a control. While pretreating cells with CORMs did not prevent the drug's overall toxicity, it effectively combated oxidative stress. Both CORM-2 and CORM-3 significantly reduced destructive oxygen free radicals induced by doxorubicin. Surprisingly, the inactive, gas-free molecules were equally effective at this. The researchers also assessed programmed cell death by measuring caspase activity. Active CORM-3 inhibited this process, showing a protective effect, whereas CORM-2 unexpectedly accelerated cell death. Additionally, CORM-3 reduced the activity of certain oxidative stress response genes without affecting inflammatory genes or final defensive protein levels. This research highlights the complex relationship between carbon monoxide releasers and chemotherapy. Although these molecules are not a cure-all for doxorubicin's toxicity, they possess valuable antioxidant properties. Crucially, the study reveals that the positive intracellular effects often stemmed from the molecule’s structural backbone itself rather than the released carbon monoxide. This discovery is a significant step toward understanding cellular mechanisms and designing better protective therapies for cancer patients. Dane dotyczą wyników z oznaczeń: cytotoksyczności, reaktywnych form tlenu, apoptozy, ekspresji genów i białek zaangażowanych w stan zapalny i stres oksydacyjny. Tue, 10 Dec 2024 00:00:00 GMT http://hdl.handle.net/11089/58757 2024-12-10T00:00:00Z