OBJECTIVES: This study aims to identify factors possibly contributing to complications in children with acute leukaemia. Despite diverse etiological causes, similar processes trigger the process of cell malignancy. Genomic instability has received considerable attention in this context. METHOD: We conducted chromosomal analysis of bone marrow cells and measured the micronuclei (Mn) level in buccal cells over time. Statistical reliability assessment was performed using Analysis of variance (ANOVA), and the data were analyzed and visualized using the SPSS 12 statistical analysis software package. RESULTS: On the 15th day of treatment, our findings confirmed a statistically significant correlation (χ2=3.88, P=0.04) between the number of blasts in the bone marrow and unfavourable outcome in patients with a near-tetraploid chromosome clone. Additionally, on the 33rd day of treatment, we observed a correlation between an elevated number of Mn and relapses. DISCUSSION: While it is commonly believed that a hyperdiploid clone with >50 chromosomes in childhood acute lymphoblastic leukaemia confers favorable outcome, our study revealed partially heterogeneous results and poor prognosis in patients with a near-tetraploid clone. We have also identified a correlation between the Mn level on the 33rd day of treatment and the development of complications. It is possible that the increased Mn values and the occurrence of relapses were influenced by the individual patient's sensitivity to the genotoxic effect of the medication.

• MeSH
• Precursor Cell Lymphoblastic Leukemia-Lymphoma * genetics   MeSH
• Bone Marrow Cells pathology   MeSH
• Child MeSH
• Humans MeSH
• Micronucleus Tests MeSH
• Micronuclei, Chromosome-Defective * MeSH
• Adolescent MeSH
• Child, Preschool MeSH
• Prognosis MeSH
• Tetraploidy MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Dle současných doporučených kritérií pro diagnostiku a staging nemoci s Lewy tělísky je nezbytné hodnocení přítomnosti Lewyho tělísek a Lewyho neuritů v konkrétních oblastech mozku. Nejpoužívanější systémy pro staging určují míru postižení na podkladě přítomnosti Lewyho patologie v konkrétních anatomických oblastech mozku v kaudo-rostrálním směru. Výběr správného klonu protilátky a využití optimalizovaného protokolu včetně efektivního odmaskování epitopu je pro vizualizaci diagnostických depozit nezbytné. Cílem naší studie bylo zhodnocení využitelnosti některých komerčně dostupných a široce používaných primárních protilátek proti alfa-synukleinu v kontextu post mortem diagnostiky a stagingu nemoci s Lewyho tělísky. Zaměřili jsme se na imunohistochemický a imunofluorescenční průkaz Lewyho patologie pomocí protilátek proti všem základním částem polypeptidového řetězce alfa-synuklein, včetně konformačně specifického klonu a klonu pro detekci posttranslačně modifikovaného proteinu.

According to the current recommended criteria for the diagnosis and staging of the Lewy body disease, it is necessary to evaluate the presence of Lewy bodies and Lewy neurites in specific areas of the brain. The most widely used staging systems assess the degree of neurodegeneration based on the distribution of Lewy pathology across specific anatomical regions of the brain, progressing in a caudo-rostral trajectory. Choosing the right antibody clone and using an optimized protocol including effective antigen retrieval is essential for the visualization of diagnostic deposits. The aim of our study was to evaluate the utility of some commercially available and widely used primary antibodies against alpha-synuclein in the context of post mortem diagnosis and staging of Lewy body disease. We focused on immunohistochemical and immunofluorescence analysis of Lewy pathology using antibodies with epitopes in all parts of the alphasynuclein polypeptide chain, including a conformation-specific clone and a clone for the detection of post-translationally modified protein.

Chronická spontánní kopřivka je charakterizována výsevem pomfů a/nebo angioedému trvajícím déle než 6 týdnů, výsevy mohou přetrvávat měsíce i roky. Toto onemocnění může vést k výraznému zhoršení kvality života. Dle současných mezinárodních doporučení jsou lékem první volby nesedativní H1 antihistaminika II. generace. Při malém terapeutickém efektu standardní dávky H1 antihistaminik II. generace je doporučeno navýšení až na čtyřnásobek této dávky. V případě nedostatečného efektu terapie antihistaminiky je indikována biologická terapie omalizumabem.

Chronic spontaneous urticaria is characterized by reccurent wheals lasting more than 6 weeks, it can persist for months or years. The disease can lead to a significant worsening in the quality of life. According to recent guidelines second-generation non-sedative antihistamines are the first-line treatment. If the therapeutic effect of the standard dose of antihistamines is insufficient, it is recommended to up-dose to 4fold. In case of insufficient effect of antihistamine therapy, biological therapy with omalizumab is indicated.

• MeSH
• Histamine Antagonists pharmacology   therapeutic use   MeSH
• Chronic Urticaria * diagnosis   therapy   MeSH
• Diagnosis, Differential MeSH
• Humans MeSH
• Omalizumab pharmacology   therapeutic use   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Review MeSH

The sodium/calcium exchanger (NCX) type 1 has been well described in various cancers, but little is known about the other two NCX types (NCX2 and NCX3). In this study, we used the selective blocker of NCX3 - YM-244769 to investigate changes in apoptosis induction, migration, proliferation, intracellular calcium and ATP in four cancer cell lines - DLD1, HeLa, MDA-MB-231 and JIMT1. In all four cell lines we observed a concentration-dependent increase in the number of apoptotic cells, as well as reduced migration and proliferation. Induction of hypoxic conditions did not alter the response of these cells to YM-244769 in any of the above-mentioned parameters. These results indicate the role of NCX3 in cancer cell migration, proliferation and apoptosis, as inhibition of NCX1 by the specific blocker SEA0400 had no significant effect on these parameters. However, we verified the effect of NCX3 inhibition by using CRISPR/Cas9 to generate clones in which the SLC8A3 (NCX3) gene was deleted, and we obtained the same results. In addition, mitochondrial respiration was impaired in the clones with NCX3 knocked-out, suggesting that NCX3 also play a role in bioenergetics. In conclusion, we have clearly shown that NCX3 plays an important anti-apoptotic, pro-migratory and proliferative role in the cancer cells by affecting mitochondrial bioenergetics, thus supporting their survival and fate.

• MeSH
• Apoptosis drug effects   MeSH
• Humans MeSH
• Mitochondria metabolism   drug effects   MeSH
• Cell Line, Tumor MeSH
• Neoplasms * metabolism   pathology   genetics   MeSH
• Cell Movement drug effects   MeSH
• Cell Proliferation drug effects   MeSH
• Sodium-Calcium Exchanger * metabolism   genetics   antagonists & inhibitors   MeSH
• Calcium metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

The parasitic protozoan Entamoeba histolytica secretes extracellular vesicles (EVs), but so far little is known about their function in the interaction with the host immune system. Infection with E. histolytica trophozoites can lead to formation of amebic liver abscesses (ALAs), in which pro-inflammatory immune responses of Ly6Chi monocytes contribute to liver damage. Men exhibit a more severe pathology as the result of higher monocyte recruitment and a stronger immune response. To investigate the role of EVs and pathogenicity in the host immune response, we studied the effect of EVs secreted by low pathogenic EhA1 and highly pathogenic EhB2 amebae on monocytes. Size and quantity of isolated EVs from both clones were similar. However, they differed in their proteome and miRNA cargo, providing insight into factors potentially involved in amebic pathogenicity. In addition, EVs were enriched in proteins with signaling peptides compared with the total protein content of trophozoites. Exposure to EVs from both clones induced monocyte activation and a pro-inflammatory immune response as evidenced by increased surface presentation of the activation marker CD38 and upregulated gene expression of key signaling pathways (including NF-κB, IL-17 and TNF signaling). The release of pro-inflammatory cytokines was increased in EV-stimulated monocytes and more so in male- than in female-derived cells. While EhA1 EV stimulation caused elevated myeloperoxidase (MPO) release by both monocytes and neutrophils, EhB2 EV stimulation did not, indicating the protective role of MPO during amebiasis. Collectively, our results suggest that parasite-released EVs contribute to the male-biased immunopathology mediated by pro-inflammatory monocytes during ALA formation.

• MeSH
• Liver Abscess, Amebic immunology   parasitology   MeSH
• Cytokines metabolism   MeSH
• Entamoebiasis immunology   parasitology   MeSH
• Entamoeba histolytica * immunology   pathogenicity   genetics   MeSH
• Extracellular Vesicles * immunology   metabolism   MeSH
• Humans MeSH
• Monocytes * immunology   parasitology   MeSH
• Signal Transduction * MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Tobacco smoke, alone or combined with alcohol, is the predominant cause of head and neck cancer (HNC). We explore how tobacco exposure contributes to cancer development by mutational signature analysis of 265 whole-genome sequenced HNC samples from eight countries. Six tobacco-associated mutational signatures were detected, including some not previously reported. Differences in HNC incidence between countries corresponded with differences in mutation burdens of tobacco-associated signatures, consistent with the dominant role of tobacco in HNC causation. Differences were found in the burden of tobacco-associated signatures between anatomical subsites, suggesting that tissue-specific factors modulate mutagenesis. We identified an association between tobacco smoking and alcohol-related signatures, indicating a combined effect of these exposures. Tobacco smoking was associated with differences in the mutational spectra, repertoire of driver mutations in cancer genes and patterns of copy number change. Our results demonstrate the multiple pathways by which tobacco smoke can influence the evolution of cancer cell clones.

• MeSH
• Tobacco Smoking * adverse effects   MeSH
• Smoking adverse effects   MeSH
• Humans MeSH
• Mutation MeSH
• Mutagenesis * genetics   MeSH
• Head and Neck Neoplasms * genetics   etiology   epidemiology   MeSH
• Whole Genome Sequencing MeSH
• DNA Copy Number Variations MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

INTRODUCTION: A critical step preceding the potential biomedical application of nanoparticles is the evaluation of their immunomodulatory effects. Such nanoparticles are expected to enter the bloodstream where they can be recognized and processed by circulating monocytes. Despite the required biocompatibility, this interaction can affect intracellular homeostasis and modulate physiological functions, particularly inflammation. This study focuses on titanium dioxide (TiO2) as an example of relatively low cytotoxic nanoparticles with potential biomedical use and aims to evaluate their possible modulatory effects on the inflammasome-based response in human primary monocytes. METHODS: Monocyte viability, phenotypic changes, and cytokine production were determined after exposure to TiO2 (diameter, 25 nm; P25) alone. In the case of the modulatory effects, we focused on NLRP3 activation. The production of IL-1β and IL-10 was evaluated after (a) simultaneous activation of monocytes with bacterial stimuli muramyl dipeptide (MDP), or lipopolysaccharide (LPS), and TiO2 (co-exposure model), (b) prior activation with TiO2 alone and subsequent exposure to bacterial stimuli MDP or LPS. The differentiation of TiO2-treated monocytes into macrophages and their polarization were also assessed. RESULTS: The selected TiO2 concentration range (30-120 μg/mL) did not induce any significant cytotoxic effects. The highest dose of TiO2 promoted monocyte survival and differentiation into macrophages, with the M2 subset being the most prevalent. Nanoparticles alone did not induce substantial production of inflammatory cytokines IL-1β, IL-6, or TNF-α. The immunomodulatory effect on NLRP3 depended on the type of costimulant used. While co-exposure of monocytes to MDP and TiO2 boosted NLRP3 activity, co-exposure to LPS and TiO2 inhibited NLRP3 by enhancing IL-10 release. The inhibitory effect of TiO2 on NLRP3 based on the promotion of IL-10 was confirmed in a post-exposure model for both costimulants. CONCLUSION: This study confirmed a non-negligible modulatory effect on primary monocytes in their inflammasome-based response and differentiation ability.

• MeSH
• Acetylmuramyl-Alanyl-Isoglutamine pharmacology   MeSH
• Cell Differentiation drug effects   MeSH
• Cytokines metabolism   MeSH
• Inflammasomes drug effects   MeSH
• Interleukin-10 metabolism   MeSH
• Interleukin-1beta metabolism   MeSH
• Metal Nanoparticles chemistry   toxicity   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Lipopolysaccharides * pharmacology   MeSH
• Macrophages drug effects   MeSH
• Monocytes * drug effects   MeSH
• Nanoparticles chemistry   toxicity   MeSH
• NLR Family, Pyrin Domain-Containing 3 Protein * metabolism   MeSH
• Titanium * chemistry   pharmacology   toxicity   MeSH
• Cell Survival * drug effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

PURPOSE: This study aimed to evaluate early-phase safety of subretinal application of AAVanc80.CAG.USH1Ca1 (OT_USH_101) in wild-type (WT) pigs, examining the effects of a vehicle control, low dose, and high dose. METHODS: Twelve WT pigs (24 eyes) were divided into three groups: four pigs each received bilateral subretinal injections of either vehicle, low dose (3.3 × 1010 vector genomes [vg] per eye), or high dose (1.0 × 1011 vg per eye). Total retinal thickness (TRT) was evaluated using optical coherence tomography and retinal function was assessed with full-field electroretinography (ff-ERG) at baseline and two months post-surgery. After necropsy, retinal changes were examined through histopathology, and human USH1C_a1/harmonin expression was assessed by quantitative PCR (qPCR) and Western blotting. RESULTS: OT_USH_101 led to high USH1C_a1 expression in WT pig retinas without significant TRT changes two months after subretinal injection. The qPCR revealed expression of the human USH1C_a1 transgene delivered by the adeno-associated virus vector. TRT changes were minimal across groups: vehicle (256 ± 21 to 243 ± 18 μm; P = 0.108), low dose (251 ± 32 to 258 ± 30 μm; P = 0.076), and high dose (242 ± 24 to 259 ± 28 μm; P = 0.590). The ff-ERG showed no significant changes in rod or cone responses. Histopathology indicated no severe retinal adverse effects in the vehicle and low dose groups. CONCLUSIONS: Early-phase clinical imaging, electrophysiology, and histopathological assessments indicated that subretinal administration of OT_USH_101 was well tolerated in the low-dose treatment arm. OT_USH_101 treatment resulted in high expression of human USH1C_a1. Although histopathological changes were not severe, more frequent changes were observed in the high-dose group.

• MeSH
• Cytoskeletal Proteins genetics   MeSH
• Dependovirus genetics   MeSH
• Electroretinography * MeSH
• Genetic Therapy methods   MeSH
• Genetic Vectors * MeSH
• Injections, Intraocular * MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Humans MeSH
• Disease Models, Animal MeSH
• Tomography, Optical Coherence * MeSH
• Swine MeSH
• Cell Cycle Proteins genetics   MeSH
• Gene Expression Regulation MeSH
• Retina * metabolism   pathology   MeSH
• Transgenes * MeSH
• Blotting, Western MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Neurons in the CNS lose regenerative potential with maturity, leading to minimal corticospinal tract (CST) axon regrowth after spinal cord injury (SCI). In young rodents, knockdown of PTEN, which antagonizes PI3K signaling by hydrolyzing PIP3, promotes axon regeneration following SCI. However, this effect diminishes in adults, potentially due to lower PI3K activation leading to reduced PIP3. This study explores whether increased PIP3 generation can promote long-distance regeneration in adults. We used a hyperactive PI3K, PI3Kδ (PIK3CD), to boost PIP3 levels in mature cortical neurons and assessed CST regeneration after SCI. Adult rats received AAV1-PIK3CD and AAV1-eGFP, or AAV1-eGFP alone, in the sensorimotor cortex concurrent with a C4 dorsal SCI. Transduced neurons showed increased pS6 levels, indicating elevated PI3K/Akt/mTOR signaling. CST regeneration, confirmed with retrograde tracing, was evaluated up to 16 weeks post injury. At 12 weeks, ∼100 axons were present at lesion sites, doubling to 200 by 16 weeks, with regeneration indices of 0.1 and 0.2, respectively. Behavioral tests showed significant improvements in paw reaching, grip strength, and ladder-rung walking in PIK3CD-treated rats, corroborated by electrophysiological recordings of cord dorsum potentials and distal flexor muscle electromyography. Thus, PI3Kδ upregulation in adult cortical neurons enhances axonal regeneration and functional recovery post SCI.

• MeSH
• Axons metabolism   physiology   MeSH
• Dependovirus genetics   MeSH
• Class I Phosphatidylinositol 3-Kinases metabolism   genetics   MeSH
• Phosphatidylinositol 3-Kinases metabolism   MeSH
• Genetic Vectors genetics   MeSH
• Rats MeSH
• Disease Models, Animal MeSH
• Neurons metabolism   MeSH
• Recovery of Function MeSH
• Spinal Cord Injuries * metabolism   therapy   genetics   MeSH
• Pyramidal Tracts * metabolism   MeSH
• Nerve Regeneration * MeSH
• Signal Transduction MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

In intravenous immunoglobulins (IVIG), and some other immunoglobulin products, protein particles have been implicated in adverse events. Role and mechanisms of immunoglobulin particles in vascular adverse effects of blood components and manufactured biologics have not been elucidated. We have developed a model of spherical silica microparticles (SiMPs) of distinct sizes 200-2000 nm coated with different IVIG- or albumin (HSA)-coronas and investigated their effects on cultured human umbilical vein endothelial cells (HUVEC). IVIG products (1-20 mg/mL), bare SiMPs or SiMPs with IVIG-corona, did not display significant toxicity to unstimulated HUVEC. In contrast, in TNFα-stimulated HUVEC, IVIG-SiMPs induced decrease of HUVEC viability compared to HSA-SiMPs, while no toxicity of soluble IVIG was observed. 200 nm IVIG-SiMPs after 24 h treatment further increased ICAM1 (intercellular adhesion molecule 1) and tissue factor surface expression, apoptosis, mammalian target of rapamacin (mTOR)-dependent activation of autophagy, and release of extracellular vesicles, positive for mitophagy markers. Toxic effects of IVIG-SiMPs were most prominent for 200 nm SiMPs and decreased with larger SiMP size. Using blocking antibodies, toxicity of IVIG-SiMPs was found dependent on FcγRII receptor expression on HUVEC, which increased after TNFα-stimulation. Similar results were observed with different IVIG products and research grade IgG preparations. In conclusion, submicron particles with immunoglobulin corona induced size-dependent toxicity in TNFα-stimulated HUVEC via FcγRII receptors, associated with apoptosis and mTOR-dependent activation of autophagy. Testing of IVIG toxicity in endothelial cells prestimulated with proinflammatory cytokines is relevant to clinical conditions. Our results warrant further studies on endothelial toxicity of sub-visible immunoglobulin particles.

• MeSH
• Apoptosis drug effects   MeSH
• Autophagy * drug effects   MeSH
• Human Umbilical Vein Endothelial Cells * drug effects   metabolism   MeSH
• Immunoglobulins, Intravenous * MeSH
• Humans MeSH
• Intercellular Adhesion Molecule-1 metabolism   MeSH
• Silicon Dioxide chemistry   toxicity   MeSH
• Protein Corona metabolism   MeSH
• Receptors, IgG * metabolism   MeSH
• Tumor Necrosis Factor-alpha * metabolism   MeSH
• TOR Serine-Threonine Kinases metabolism   MeSH
• Particle Size MeSH
• Cell Survival drug effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH