No abstract

The recent JAK1/2 inhibitor trial in myeloproliferative neoplasms (MPNs) showed that reducing inflammation can be more beneficial than targeting gene mutants. We evaluated the proinflammatory IL-6 cytokine and JAK-STAT signaling pathway related genes in circulating CD34+ cells of MPNs. Regarding laboratory data, leukocytosis has been observed in polycythemia vera (PV) and JAK2V617F mutation positive versus negative primary myelofibrosis (PMF) patients. Moreover, thrombocytosis was reduced by JAK2V617F allele burden in essential thrombocythemia (ET) and PMF. 261 significantly changed genes have been detected in PV, 82 in ET, and 94 genes in PMF. The following JAK-STAT signaling pathway related genes had augmented expression in CD34+ cells of MPNs: CCND3 and IL23A regardless of JAK2V617F allele burden; CSF3R, IL6ST, and STAT1/2 in ET and PV with JAK2V617F mutation; and AKT2, IFNGR2, PIM1, PTPN11, and STAT3 only in PV. STAT5A gene expression was generally reduced in MPNs. IL-6 cytokine levels were increased in plasma, as well as IL-6 protein levels in bone marrow stroma of MPNs, dependent on JAK2V617F mutation presence in ET and PMF patients. Therefore, the JAK2V617F mutant allele burden participated in inflammation biomarkers induction and related signaling pathways activation in MPNs.

From our data we conclude that the S100A8 and S100A9 granulocyte and plasma levels are increased in MPN patients, along with inflammation markers, depending on their JAK2V617F mutation allele burden. We also found that S100A8/9-mediated inhibition of the proliferation-related AKT and ERK1/2 signaling pathways can be decreased by CALR mutation-dependent TLR4 blocking and increased by RAGE inhibition in MPN.

Aim To examine the potential systemic toxicity of nanostructured materials based on calcium silicate and calcium aluminate, for potential application in Dentistry. Methodology Twenty‐four Albino Wistar rats aged 2 months were used as an in vivo animal model for subcutaneous implantation of the investigated materials, placed in polyethylene tubes. Thirty days after implantation, the livers of the rats were analysed and following histological and stereological parameters were evaluated for volume density of hepatocytes and blood sinusoids, number and numerical density of hepatocytes, surface of hepatocytes and their nucleuses, nucleocytoplasmic ratio and mitotic index of hepatocytes. Stereological measurements were achieved using Cavalieri's principle, with grid P2 and unbiased analysis. Additionally, immunohistochemistry studies were performed to further analyse changes in liver tissue. Several haematological and biochemical parameters of blood of experimental animals were also analysed, as well as local tissue reactions around the implants. Statistical analysis was performed using parametric (anova and t‐test) and nonparametric tests (Kruskal–Wallis and Mann–Whitney U‐test) depending on data distribution. Results Implanted dental cements led to an increase in stereological and histological parameters in liver tissue compared to control rats. Although the investigated parameters mostly showed significant differences between control and experimental animals, the liver tissue of the experimental animals did not have visible signs of pathological changes. This was supported by the analysis of blood parameters which were not significantly different between control and experimental animals. Also, the subcutaneous tissues had minimal inflammatory reactions. Immunohistochemistry studies revealed that nanostructured materials induced proliferation of hepatocytes, but that the immunological response to the materials was not strong enough to induce proliferation of immunoreactive cells in liver in the observed time period. Conclusions This study was performed as a contribution to the attestation of the biocompatibility of dental cements based on calcium silicate and calcium aluminate. Although these materials induced several changes in the liver structure, they were not clinically relevant and represent a normal and reversible response of the liver to the presence of biocompatible materials in the body. Blood and immunohistochemistry analyses and local tissue reactions further confirmed that these materials possess good biocompatible potential.

Increased angiogenesis in BCR-ABL1 negative myeloproliferative neoplasms (MPNs) has been recognized, but its connection with clinical and molecular markers needs to be defined. The aims of study were to (1) assess bone marrow (BM) angiogenesis measured by microvessel density (MVD) using CD34 and CD105 antibodies; (2) analyze correlation of MVD with plasma angiogenic factors including vascular endothelial growth factor, basic fibroblast growth factor, and interleukin-8; (3) examine the association of MVD with clinicopathological and molecular markers. We examined 90 de novo MPN patients (30 polycythemia vera (PV), primary myelofibrosis (PMF), essential thrombocythemia (ET)) and 10 age-matched controls. MVD was analyzed by immunohistochemistry "hot spot" method, angiogenic factors by immunoassay and JAK2V617F, and CALR mutations by DNA sequencing and allelic PCR. MVD was significantly increased in MPNs compared to controls (PMF> PV > ET). Correlation between MVD and plasma angiogenic factors was found in MPNs. MVD was significantly increased in patients with JAK2V617F mutation and correlated with JAK2 mutant allele burden (CD34-MVD: ρ = 0.491, p < 0.001; CD105-MVD: ρ = 0.276, p = 0.02) but not with CALR mutation. MVD correlated with leukocyte count, serum lactate dehydrogenase, hepatomegaly, and splenomegaly. BM fibrosis was significantly associated with CD34-MVD, CD105-MVD, interleukin-8, and JAK2 mutant allele burden. JAK2 homozygote status had positive predictive value (100%) for BM fibrosis. Patients with prefibrotic PMF had significantly higher MVD than patients with ET, and we could recommend MVD to be additional histopathological marker to distinguish these two entities. This study also highlights the strong correlation of MVD with plasma angiogenic factors, JAK2 mutant allele burden, and BM fibrosis in MPNs.