ELISA-like Analysis of Cisplatinated DNA Using Magnetic Separation

Cisplatin belongs to the most widely used cytostatic drugs. The determination of the presence of the DNA-cisplatin adducts may not only signal the guanine-rich regions but also monitor the interaction reaction between DNA and the drug in terms of speed of interaction. In this work, the combined advantages of magnetic particles-based isolation/purification with fluorescent properties of quantum dots (QDs) and antibodies targeted on specific recognition of DNA-cisplatin adducts are demonstrated. The formation of a complex between magnetic particles with surface modified by anti-dsDNA antibody, cisplatin-modified DNA and QDs labelled anti-cisplatin-modified DNA antibody was suggested and optimized

Central Disorders of Hypersomnolence: Association with Fatigue, Depression and Sleep Inertia Prevailing in Women

Fatigue, depression, and sleep inertia are frequently underdiagnosed manifestations in narcolepsy and idiopathic hypersomnia. Our cross-sectional study design included diagnostic interview accompanied by assessment instruments and aimed to explore how these factors influence disease severity as well as to elucidate any sex predisposition. One hundred and forty-eight subjects (female 63%) were divided into narcolepsy type 1 (NT1; n = 87, female = 61%), narcolepsy type 2 (NT2; n = 22, female = 59%), and idiopathic hypersomnia (IH; n = 39, female = 69%). All subjects completed a set of questionnaires: Epworth Sleepiness Scale (ESS), Hospital Anxiety and Depression Scales (HADS), Fatigue Severity Scale (FSS), and Sleep Inertia Questionnaire (SIQ). In narcoleptic subjects, questionnaire data were correlated with the Narcolepsy Severity Scale (NSS), and in subjects with idiopathic hypersomnia, with the Idiopathic Hypersomnia Severity Scale (IHSS). The highest correlation in narcoleptic subjects was found between NSS and ESS (r = 0.658; p p p p p = 0.0005), and HADS anxiety scale (r = 0.528; p p p p p p = 0.057). Our study illustrates that more attention should be focused on pathophysiological mechanisms and associations of fatigue, depression, as well as sleep inertia in these diseases; they influence the course of both illnesses, particularly in women

A Simple RNA Target Capture NGS Strategy for Fusion Genes Assessment in the Diagnostics of Pediatric B-cell Acute Lymphoblastic Leukemia

Acute lymphoblastic leukemia (ALL) is the most frequent pediatric cancer. Fusion genes are hallmarks of ALL, and they are used as biomarkers for risk stratification as well as targets for precision medicine. Hence, clinical diagnostics pursues broad and comprehensive strategies for accurate discovery of fusion genes. Currently, the gold standard methodologies for fusion gene detection are fluorescence in situ hybridization and polymerase chain reaction; these, however, lack sensitivity for the identification of new fusion genes and breakpoints. In this study, we implemented a simple operating procedure (OP) for detecting fusion genes. The OP employs RNA CaptureSeq, a versatile and effortless next-generation sequencing assay, and an in-house as well as a purpose-built bioinformatics pipeline for the subsequent data analysis. The OP was evaluated on a cohort of 89 B-cell precursor ALL (BCP-ALL) pediatric samples annotated as negative for fusion genes by the standard techniques. The OP confirmed 51 samples as negative for fusion genes, and, more importantly, it identified known (KMT2A rearrangements) as well as new fusion events (JAK2 rearrangements) in the remaining 38 investigated samples, of which 16 fusion genes had prognostic significance. Herein, we describe the OP and its deployment into routine ALL diagnostics, which will allow substantial improvements in both patient risk stratification and precision medicine

Selected haematological parameters of erythrocytes mixture with platelets and oxidative stress (GSH/GSSG) in the mixture after application of Pt derivatives encapsulated in liposomes or alone in concentrations 0, 12.5, 25, 50, 100 and 200 μg/mL of Pt and 0, 0.6, 1.3, 2.5, 5 and 10 mg/mL of liposomes.

<p>(A) Histograms of (a) erythrocytes and (b) platelets, and (c) visualisations of liposomes in the Baso channel after application of LipoCisPt, CisPt, LipoPtNPs and PtNPs. GSH/GSSG ratio of erythrocytes mixture with platelets in the same concentrations as in A—(B) LipoCisPt, (C) CisPt, (D) LipoPtNPs, and (E) PtNPs. (F) Mean of GSH/GSSG ratio. For all measurement n = 3, significant difference is indicated by *p<0.05.</p

The effect of platinum derivatives on both bacterial and human cells after application of platinum derivatives alone or encapsulated in liposomes.

<p>Relative change of growth rate of (A) <i>S</i>. <i>aureus</i> bacterial culture and (C) HFF after treatment with Pt-derivatives (100 μg/mL of Pt) during 24 h (bacterial culture) and 100 h (HFF) long experiments. (a) Cell culture without any treatment, (b) LipoCisPt, (c) CisPt, (d) LipoPtNPs and (e) PtNPs. Percentage survival of (B) <i>S</i>. <i>aureus</i> bacterial culture and (D) human foreskin fibroblasts (HFF) after 24 and 100 h of treatment with Pt-derivatives (100 μg/mL of Pt) alone and encapsulated in liposomes. For all measurement n = 3.</p