Fast and efficient homing and engraftment of hematopoietic stem progenitor cells (HSPCs) is crucial for positive clinical outcomes from transplantation. We found that this process depends on activation of the Nlrp3 inflammasome, both in the HSPCs to be transplanted and in the cells in the recipient bone marrow (BM) microenvironment. For the first time we provide evidence that functional deficiency in the Nlrp3 inflammasome in transplanted cells or in the host microenvironment leads to defective homing and engraftment. At the molecular level, functional deficiency of the Nlrp3 inflammasome in HSPCs leads to their defective migration in response to the major BM homing chemoattractant stromal-derived factor 1 (SDF-1) and to other supportive chemoattractants, including sphingosine-1-phosphate (S1P) and extracellular adenosine triphosphate (eATP). We report that activation of the Nlrp3 inflammasome increases autocrine release of eATP, which promotes incorporation of the CXCR4 receptor into membrane lipid rafts at the leading surface of migrating cells. On the other hand, a lack of Nlrp3 inflammasome expression in BM conditioned for transplantation leads to a decrease in expression of SDF-1 and danger-associated molecular pattern molecules (DAMPs), which are responsible for activation of the complement cascade (ComC), which in turn facilitates the homing and engraftment of HSPCs.

An efficient harvest of hematopoietic stem/progenitor cells (HSPCs) after pharmacological mobilization from the bone marrow (BM) into peripheral blood (PB) and subsequent proper homing and engraftment of these cells are crucial for clinical outcomes from hematopoietic transplants. Since extracellular adenosine triphosphate (eATP) plays an important role in both processes as an activator of sterile inflammation in the bone marrow microenvironment, we focused on the role of Pannexin-1 channel in the secretion of ATP to trigger both egress of HSPCs out of BM into PB as well as in reverse process that is their homing to BM niches after transplantation into myeloablated recipient. We employed a specific blocking peptide against Pannexin-1 channel and noticed decreased mobilization efficiency of HSPCs as well as other types of BM-residing stem cells including mesenchymal stroma cells (MSCs), endothelial progenitors (EPCs), and very small embryonic-like stem cells (VSELs). To explain better a role of Pannexin-1, we report that eATP activated Nlrp3 inflammasome in Gr-1⁺ and CD11b⁺ cells enriched for granulocytes and monocytes. This led to release of danger-associated molecular pattern molecules (DAMPs) and mitochondrial DNA (miDNA) that activate complement cascade (ComC) required for optimal egress of HSPCs from BM. On the other hand, Pannexin-1 channel blockage in transplant recipient mice leads to a defect in homing and engraftment of HSPCs. Based on this, Pannexin-1 channel as a source of eATP plays an important role in HSPCs trafficking.

Introduction:The purpose of the study was to determine and model the growth rates ofL. monocytogenesin cooked cured ham stored at various temperatures.Material and Methods:Samples of cured ham were artificially contaminated with a mixture of threeL. monocytogenesstrains and stored at 3, 6, 9, 12, or 15°C for 16 days. The number of listeriae was determined after 0, 1, 2, 3, 5, 7, 9, 12, 14, and 16 days. A series of decimal dilutions were prepared from each sample and plated onto ALOA agar, after which the plates were incubated at 37°C for 48 h under aerobic conditions. The bacterial counts were logarithmised and analysed statistically. Five repetitions of the experiment were performed.Results:Both storage temperature and time were found to significantly influence the growth rate of listeriae (P < 0.01). The test bacteria growth curves were fitted to three primary models: the Gompertz, Baranyi, and logistic. The mean square error (MSE) and Akaike’s information criterion (AIC) were calculated to evaluate the goodness of fit. It transpired that the logistic model fit the experimental data best. The natural logarithms ofL. monocytogenes’mean growth rates from this model were fitted to two secondary models: the square root and polynomial.Conclusion:Modelling in both secondary types can predict the growth rates ofL. monocytogenesin cooked cured ham stored at each studied temperature, but mathematical validation showed the polynomial model to be more accurate.

We found that circadian changes in ATP level in peripheral blood (PB) activate the Nlrp3 inflammasome, which triggers diurnal release of hematopoietic stem/progenitor cells (HSPCs) from murine bone marrow (BM) into PB. Consistent with this finding, we observed circadian changes in expression of mRNA for Nlrp3 inflammasome-related genes, including Nlrp3, caspase 1, IL-1β, IL-18, gasdermin (GSDMD), HMGB1, and S100A9. Circadian release of HSPCs from BM into PB as well as expression of Nlrp3-associated genes was decreased in mice in which pannexin 1-mediated secretion of ATP was inhibited by the blocking peptide 10Panx and in animals exposed to the specific small-molecule inhibitor of the Nlrp3 inflammasome MCC950. In addition to HSPCs, a similar decrease in diurnal cell counts was observed for mesenchymal stromal cells (MSCs), endothelial progenitor cells (EPCs), and very small embryonic-like stem cells (VSELs). These results shed more light on the complexity of circadian regulation of HSPC release into PB, which is coordinated in a purinergic signaling-, innate immunity-dependent manner. Moreover, in addition to circadian changes in expression of the Nlrp3 inflammasome we also observed diurnal changes in expression of other inflammasomes, including Aim2, Nrp1a, and Nlrp1b.

Campylobacter jejuni inactivation by high pressure processing (HPP) in poultry meat (chicken breast) was investigated. The pressure was created by high hydrostatic pressure piston-cylinder food processor. Contaminated with C. jejuni (108 CFU g-1) samples of ground poultry meat were hermetically sealed in a polyamide-polyethylene bags and exposed to HPP for 9 different combinations of pressure (200 MPa, 300 MPa and 400 MPa) and time (5 min, 10 min and 15 min). Quantitative bacteriological analysis was carried out in order to determine the number of surviving C. jejuni cells. The obtained results showed that C. jejuni is relatively sensitive to high pressure treatment as compared to other food-borne pathogens. The loss of C. jejuni viability increased in a dose- and time-dependent manner. On the basis of the results, D-values were calculated. For reduction C. jejuni in poultry meat by 6 log units (6D-values), considered as sufficient for consumer protection, the application of 300 MPa for 8.73 min, or 400 MPa for 4.37 min is needed. The linear regression equations, which has been calculated on the basis of this study, allows to determine the degree of C. jejuni reduction in poultry meat for any selected duration of pressurization in a given pressure range.